Substituted 3-haloallylamine inhibitors of SSAO and uses thereof

ABSTRACT

The invention also relates to methods of using compounds of Formula I, or pharmaceutically acceptable salt or derivatives thereof, for the treatment of a variety of indications, e.g., inflammatory diseases, ocular diseases, fibrotic diseases, diabetes-induced diseases and cancer.

TECHNICAL FIELD

The present invention relates to novel compounds which are capable ofinhibiting certain amine oxidase enzymes. These compounds are useful fortreatment of a variety of indications, e.g., the symptoms ofinflammation and/or fibrosis in human subjects as well as in pets andlivestock, the treatment of psychological diseases, neurodegenerativedisorders, and the like. In addition, the present invention relates topharmaceutical compositions containing these compounds, as well asvarious uses therefore.

BACKGROUND

Semicarbazide-sensitive amine oxidase (SSAO), also known as primaryamine oxidase, plasma amine oxidase and benzylamine oxidase, isidentical in structure to vascular adhesion protein-1 (VAP-1). In thefollowing discussion, SSAO/VAP-1 is used to describe this protein. Therole of this protein in inflammatory diseases has been reviewed (see,for example, Smith D. J. and Vaino P. J., Targeting Vascular AdhesionProtein-1 to Treat Autoimmune and Inflammatory Diseases. Ann. N.Y. Acad.Sci. 2007, 1110, 382-388; and McDonald I. A. et al., SemicarbazideSensitive Amine Oxidase and Vascular Adhesion Protein-1: One ProteinBeing Validated as a Therapeutic Target for Inflammatory Diseases.Annual Reports in Medicinal Chemistry, 2008, 43, 229-241).

In most organisms, including humans, two families of mammalian amineoxidases metabolize various mono-, di-, and polyamines producedendogenously or absorbed from exogenous sources. These include themonoamine oxidases (MAO-A and MAO-B) which are present in themitochondria of most cell types and use covalently bound flavin adeninedinucleotide (FAD) as the cofactor. Polyamine oxidase is anotherFAD-dependent amine oxidase which oxidatively deaminates spermine andspermidine. SSAO/VAP-1 belongs to the second family which is dependenton copper and uses other co-factors apart from FAD, such as an oxidizedtyrosine residue (abbreviated as TPQ or LTQ). MAO and SSAO/VAP-1oxidatively deaminate some common substrates which includes themonoamines such dopamine, tyramine and benzylamine. SSAO/VAP-1 alsooxidizes endogenous methylamine and aminoacetone.

Some of these enzymes were originally defined by the ability of certaincompounds to inhibit the enzymatic activity thereof. For example MAO-Ais selectively inhibited by clorgyline, MAO-B by L-deprenyl, whileneither clorgyline nor L-deprenyl can inhibit the amine oxidase activityof SSAO/VAP-1. SSAO/VAP-1 can be inhibited by semicarbazide, hence thename semicarbazide sensitive amine oxidase.

SSAO/VAP-1 is an ectoenzyme containing a very short cytoplasmic tail, asingle transmembrane domain, and a large, highly glycosylatedextracellular domain which contains the active center for the amineoxidase activity. SSAO/VAP-1 is also present in a soluble formcirculating in the plasma of some animals. It has been shown that thisform is a cleaved product of membrane-bound SSAO/VAP-1.

SSAO/VAP-1 appears to have two physiological functions: the first is theamine oxidase activity mentioned above and the second is cell adhesionactivity. Both activities are associated with inflammatory processes.SSAO/VAP-1 was shown to play an important role in extravasation ofinflammatory cells from the circulation to sites of inflammation (SalmiM. and Jalkanen S., VAP-1: an adhesin and an enzyme. Trends Immunol.2001, 22, 211-216). VAP-1 antibodies have been demonstrated to attenuateinflammatory processes by blocking the adhesion site of the SSAO/VAP-1protein and, together with a substantial body of evidence of in vitroand in vivo knockouts, it is now clear that SSAO/VAP-1 is an importantcellular mediator of inflammation. Transgenic mice lacking SSAO/VAP-1show reduced adhesion of leukocytes to endothelial cells, reducedlymphocyte homing to the lymph nodes and a concomitant attenuatedinflammatory response in a peritonitis model. These animals wereotherwise healthy, grew normally, were fertile, and examination ofvarious organs and tissues showed the normal phenotype. Furthermore,inhibitors of the amine oxidase activity of SSAO/VAP-1 have been foundto interfere with leukocyte rolling, adhesion and extravasation and,similar to SSAO/VAP-1 antibodies, exhibit anti-inflammatory properties.

Inflammation is the first response of the immune system to infection orirritation. The migration of leukocytes from the circulation intotissues is essential for this process. Inappropriate inflammatoryresponses can result in local inflammation of otherwise healthy tissuewhich can lead to disorders such as rheumatoid arthritis, inflammatorybowel disease, multiple sclerosis and respiratory diseases. Leukocytesfirst adhere to the endothelium via binding to adhesion molecules beforethey can start the process of passing through the walls of the bloodvessels. Membrane bound SSAO/VAP-1 is abundantly expressed in vascularendothelial cells such as high venule endothelial cells (HVE) oflymphatic organs and is also expressed in hepatic sinusoidal endothelialcells (HSEC), smooth muscle cells and adipocytes. The expression ofSSAO/VAP-1 on the cell surface of endothelial cells is tightly regulatedand is increased during inflammation. In the presence of an SSAO/VAP-1substrate (benzylamine), NFκB was activated in HSECs together withup-regulation of other adhesion molecules, E-selectin and chemokineCXCL8 (IL-8) in vitro. A recent study confirms this result by showing(by mutagenesis) that the transcription and translation of E-selectinand P-selectin is induced by the enzyme activity of SSAO/VAP-1. Theseresults suggest an important role of the amine oxidase activity ofSSAO/VAP-1 in the inflammatory response. It has been reported that theoxidase activity of SSAO/VAP-1 induces endothelial E- and P-selectinsand leukocyte binding (Jalkanen, S. et al., The oxidase activity ofvascular adhesion protein-1 (VAP-1) induces endothelial E- andP-selectins and leukocyte binding. Blood 2007, 110, 1864-1870).

Excessive and chronic inflammatory responses have been associated withthe symptoms of many chronic diseases, such as rheumatoid arthritis,multiple sclerosis, asthma and chronic obstructive pulmonary disease(COPD). Patients suffering from either atopic eczema or psoriasis (bothchronic inflammatory skin disorders) have higher levels of SSAO/VAP-1positive cells in their skin compared to skin from healthy controls.

Asthma can be considered a disease resulting from chronic inflammationof the airways which results in bronchoconstriction and excessivebuild-up of mucus. Many patients can be adequately treated withbronchodilators (eg, β2 agonists, leukotriene antagonists and withinhaled steroids). However, up to about 20% of patients suffer fromsevere asthma and don't respond well to these treatments. A subset ofthese patients are resistant to inhaled steroids and present with highneutrophil counts in their lung fluids. SSAO/VAP-1 is expressed in thelungs and plays a role in the trafficking of neutrophils.

Another subset of asthma patients is acutely sensitive to viralinfections of the airways; such infections exacerbate the underlyinginflammation and can lead to severe asthma attacks.

It has been recently recognized that patients suffering from cysticfibrosis frequently suffer from persistent lung inflammation which canbe independent from chronic lung infection. It has been argued thattissue damage in cystic fibrosis patients is due to mediators releasedby neutrophils. While standard antibiotic treatment to clear bacterialinfection would be expected to resolve the underlying inflammation ifthe inflammation were solely due to the infection, data from recentstudies demonstrate that this is not the case and that the airways arein a neutrophil-driven pro-inflammatory state primed for excessive andprolonged inflammatory response to bacterial infection. See Rao S. andGrigg J., New insights into pulmonary inflammation in cystic fibrosis.Arch Dis Child 2006, 91:786-788.

SSAO/VAP-1 is also highly expressed in adipocytes where it plays a rolein glucose transport independent of the presence of insulin. It has beenobserved that levels of plasma SSAO/VAP-1 are increased in patientssuffering from diabetes. Elevated levels of plasma SSAO/VAP-1 have beenfound in patients suffering from other illnesses, such as congestiveheart failure and liver cirrhosis. It has been suggested that SSAO/VAP-1is associated with most, if not all, inflammatory diseases whether theinflammation is in response to an immune response or subsequent to otherevents such as occlusion and reperfusion of blood vessels.

It has been recognized in recent years that SSAO/VAP-1 is expressed insinusoidal endothelial cells in the liver and that this protein isbelieved to be associated with hepatic disease, in particular liverfibrosis (Weston C. J. and Adams D. H., Hepatic consequences of vascularadhesion protein-1 expression, J Neural Transm 2011; 118:1055-1064).Furthermore, a VAP-1 antibody and a small molecule inhibitor were foundto attenuate carbon tetrachloride induced fibrosis in mice. Thus,SSAO/VAP-1 inhibitors have the potential to treat fibrotic disease (WO2011/029996). It has been recently reported that oxidation ofmethylamine by SSAO/VAP-1 in the presence of tumor necrosis factor αinduces the expression of MAdCAM-1 in hepatic vessels, and that this isassociated with the hepatic complications of inflammatory bowel disease(IBD) (Liaskou W. et al., Regulation of Mucosal Addressin Cell AdhesionMolecule 1 Expression in Human and Mice by Vascular Adhesion Protein 1Amine Oxidase Activity, Hepatology 2011; 53, 661-672).

It has been reported that SSAO/VAP-1 inhibitors can attenuateangiogenesis and lymphangiogenesis, and that these inhibitors offerpotential to treat ocular diseases such as macular degeneration, cornealangiogenesis, cataracts, and inflammatory conditions such as uveitis (US2009/0170770; WO 2009/051223; Noda K., et al., Inhibition of vascularadhesion protein-1 suppresses endotoxin-induced uveitis, FASEB J. 2008,22, 1094-1103).

Increased levels of SSAO/VAP-1 were observed in the serum of patientssuffering from hepatocellular carcinoma. In a murine melanoma model,small molecule SSAO/VAP-1 inhibitors were shown to retard tumor growth,in contrast to VAP-1 antibodies which had no activity (Weston C. J. andAdams D. H., Hepatic consequences of vascular adhesion protein-1expression, J Neural Transm 2011, 118, 1055-1064).

It was reported that SSAO/VAP-1 plays an important role in cancerbiology (Marttila-Ichihara F. et al. Small-Molecule Inhibitors ofVascular Adhesion Protein-1 Reduce the Accumulation of Myeloid Cellsinto Tumors and Attenuate Tumor Growth in Mice. The Journal ofImmunology, 2010, 184, 3164-3173). SSAO/VAP-1 small molecule inhibitorsreduced the number of proangiogenic Gr-1+CD11b+ myeloid cells inmelanomas and lymphomas.

During the SSAO/VAP-1 amine oxidase catalytic cycle the covalently boundcofactor, TPQ, is first reduced, and then re-oxidized by oxygen in thepresence of copper with the generation of hydrogen peroxide as aby-product. It has been speculated that excessive hydrogen peroxideconcentrations can be deleterious and may contribute to the pathology ofvarious inflammatory and neurodegenerative processes (Götz M. E., etal., Oxidative stress: Free radical production in neural degeneration.Pharmacol Ther 1994, 63, 37-122).

Inflammation is believed to be an important feature of neurodegenerativediseases such as Parkinson's disease, Alzheimer's disease and multiplesclerosis, and similarly is a feature of the pathophysiology that occursafter a cerebral occlusion/reperfusion event (Aktas, O. et al., Neuronaldamage in brain inflammation. Arch Neurol 2007, 64, 185-9). Excessiveactivity SSAO/VAP-1 has been independently implicated in these processes(Xu, H-L., et al., Vascular Adhesion Protein-1 plays an important rolein postischemic inflammation and neuropathology in diabetic,estrogen-treated ovariectomized female rats subjected to transientforebrain ischemia. Journal Pharmacology and Experimental Therapeutics,2006, 317, 19-26).

Some known MAO inhibitors also inhibit SSAO/VAP-1 (e.g., the MAO-Binhibitor Mofegiline illustrated below). Mofegiline has been reported toinhibit experimental autoimmune encephalomyelitis (US 2006/0025438 A1).This inhibitor is a member of the haloallylamine family of MAOinhibitors; the halogen in Mofegiline is fluorine. Fluoroallylamineinhibitors are described in U.S. Pat. No. 4,454,158. There have beenreports of a chloroallylamine, MDL72274 (illustrated below), selectivelyinhibiting rat SSAO/VAP-1 compared to MAO-A and MAO-B:

Additional fluoroallylamine inhibitors are described in U.S. Pat. No.4,699,928; the two compounds illustrated below were described asselective inhibitors of MAO-B:

Other examples structurally related to Mofegiline can be found in WO2007/120528.

Haloallylamine compounds that differ from Mofegiline in core structurehave been synthesized and were shown to inhibit the amine oxidaseactivity from copper-dependent amine oxidases from a number of species(see Kim J., et al., Inactivation of bovine plasma amine oxidase byhaloallylamines. Bioorg Med Chem 2006, 14, 1444-1453). These compoundshave been included in a patent application (WO 2007/005737):

WO 2009/066152 describes a family of 3-substituted 3-haloallylaminesthat are inhibitors of SSAO/VAP-1 and are claimed as treatment for avariety of indications, including inflammatory disease. The followingcompounds are specifically described:

References to the effects of SSAO/VAP-1 inhibitors in various animalmodels of disease can be found in the review publication by McDonald I.A. et al., Semicarbazide Sensitive Amine Oxidase and Vascular AdhesionProtein-1: One Protein Being Validated as a Therapeutic Target forInflammatory Diseases. Annual Reports in Medicinal Chemistry, 2008, 43,229-241 and in the following publications, O'Rourke A. M. et al.,Anti-inflammatory effects of LJP 1586[Z-3-fluoro-2-(4-methoxybenzyl)allylamine hydrochloride], an amine-basedinhibitor of semicarbazide-sensitive amine oxidase activity. J.Pharmacol. Exp. Ther., 2008, 324, 867-875; and O'Rourke A. M. et al.,Benefit of inhibiting SSAO in relapsing experimental encephalomyelitis.J. Neural. Transm., 2007, 114, 845-849.

SUMMARY

The present invention provides substituted haloallylamine compounds thatinhibit SSAO/VAP-1. Surprisingly, modification of2-substituted-3-haloallylamine structures described previously has ledto the development of novel compounds that are potent inhibitors of thehuman SSAO/VAP-1 enzyme and which have much improved pharmacological andsafety properties. These compounds are very potent on SSAO/VAP-1 andwere surprisingly found to be very weak inhibitors of other familymembers, such as monoamine oxidase A, monoamine oxidase B, diamineoxidase, lysyl oxidase, and lysyl-like amine oxidases LOX 1-4.

A first aspect of the invention provides for a compound of Formula I:

or a stereoisomer, pharmaceutically acceptable salt, polymorphic form,solvate or prodrug thereof; wherein:R¹ and R⁴ are independently hydrogen or optionally substitutedC₁₋₆alkyl;R² and R³ are independently selected from the group consisting ofhydrogen, chlorine and fluorine; provided, however, that R² and R³ arenot hydrogen at the same time;R⁵ is an optionally substituted arylene group;R⁶ is selected from

R⁷ and R⁸ are independently selected from the group consisting ofhydrogen, optionally substituted C₁₋₆alkyl and optionally substitutedC₃₋₇cycloalkyl; andX is CH₂, oxygen, sulfur or SO₂.

A second aspect of the invention provides for a pharmaceuticalcomposition comprising a compound according to the first aspect of theinvention, or a pharmaceutically acceptable salt or solvate thereof, andat least one pharmaceutically acceptable excipient, carrier or diluent.

A third aspect of the invention provides for a method of inhibiting theamine oxidase activity of SSAO/VAP-1 in a subject in need thereof, saidmethod comprising administering to said subject an effective amount of acompound according to the first aspect of the invention, or apharmaceutically acceptable salt or solvate thereof, or a compositionaccording to the second aspect of the invention.

A fourth aspect of the invention provides for a method of treating adisease associated with or modulated by SSAO/VAP-1 protein, said methodcomprising administering to a subject in need thereof a therapeuticallyeffective amount of a compound according to the first aspect of theinvention, or a pharmaceutically acceptable salt or solvate thereof, ora composition according to the second aspect of the invention.

A fifth aspect of the invention provides for a method of treating adisease associated with or modulated by SSAO/VAP-1, said methodcomprising administering to a subject in need thereof a therapeuticallyeffective amount of a compound according to the first aspect of theinvention, or a pharmaceutically acceptable salt or solvate thereof, ora composition according to the second aspect of the invention.

A sixth aspect of the invention provides for use of a compound accordingto the first aspect of the invention, or a pharmaceutically acceptablesalt or solvate thereof, for the manufacture of a medicament fortreating a disease associated with or modulated by SSAO/VAP-1 protein.

A seventh aspect of the invention provides for a compound according tothe first aspect of the invention, or a pharmaceutically acceptable saltor solvate thereof, for use in treating a disease associated with ormodulated by SSAO/VAP-1 protein.

In another aspect, the present invention describes the synthesis and useof compounds which inhibit the amine oxidase activity of SSAO/VAP-1, anddescribes the use of such inhibitors to treat patients sufferinginflammatory diseases.

The compounds of the present invention are useful for the treatment ofthe symptoms of inflammation and/or fibrosis in human subjects as wellas in pets and livestock. Human inflammatory diseases contemplated fortreatment herein include arthritis, Crohn's disease, irritable boweldisease, psoriasis, eosinophilic asthma, severe asthma, virallyexacerbated asthma, chronic pulmonary obstructive disease, cysticfibrosis, bronchiectasis, atherosclerosis, inflammation due to diabetes,inflammatory cell-mediated tissue destruction following stroke, and thelike. Human fibrotic diseases and disorders contemplated for treatmentherein include idiopathic pulmonary fibrosis or other interstitial lungdiseases, liver fibrosis, kidney fibrosis, fibrosis of other organs andtissues, radiation induced fibrosis, and the like.

The compounds of the present invention are also useful for the treatmentof bacteria-induced lung inflammation associated with cystic fibrosis.Treatment can be both prophylactic and therapeutic. Furthermore, thecompounds of the present invention are useful for the treatment of otherbacteria-induced lung diseases such as sepsis, acute respiratorydistress syndrome (ARDS), acute lung injury (ALI), transfusion inducedlung injury (TRALI), and the like.

The compounds of the present invention are also useful for the treatmentof ocular diseases, such as uveitis and macular degeneration.

The compounds of the present invention are also useful as an adjuncttherapy to treat cancer. In combination with standard and novelchemotherapeutic agents, the compounds of the present invention can leadto better control of the cancer, and to help reduce metastatic secondarycancers.

Since SSAO/VAP-1 small molecule inhibitors actively attenuate neutrophillevels in the lipopolysaccharide (LPS) mouse model of lung neutrophilia,such molecules have the potential to treat steroid resistant asthma inhuman subjects. Accordingly, in accordance with one aspect of thepresent invention, there are provided methods for treating patients withan inhibitor of SSAO/VAP-1 either as a prophylactic or therapeutic agentto reduce neutrophil levels and treat the symptoms of severe asthma.

In accordance with another aspect of the present invention, there areprovided methods for treating patients with an inhibitor of SSAO/VAP-1either as a prophylactic agent or as a therapeutic agent to treaton-going disease.

In accordance with still another aspect of the present invention, thereare provided methods for the use of an SSAO/VAP-1 inhibitor to modulatethe concentration of neutrophils in the airways and to treat theunderlying cause of inflammation in patients suffering from inflammationof the airways.

In accordance with yet another aspect of the present invention, thereare provided methods for treating patients suffering from liver fibrosiswith an SSAO/VAP-1 inhibitor.

In accordance with a further aspect of the present invention, there areprovided methods for treating patients suffering from ocular diseasewith an SSAO/VAP-1 inhibitor to treat symptoms of the disease.

Since SSAO/VAP-1 is expressed in various cancer types, in accordancewith yet another aspect of the present invention, there is contemplatedthe use of SSAO/VAP-1 inhibitors as adjunctive therapy to treat patientssuffering from cancers which express SSAO/VAP-1.

In one embodiment of the methods and uses of the present invention thedisease is inflammation. In another embodiment the inflammation isassociated with liver disease. In a further embodiment the inflammationis associated with respiratory disease. In a still further embodimentthe inflammation is associated with cystic fibrosis. In anotherembodiment the inflammation is associated with asthma or chronicobstructive pulmonary disease. In a further embodiment the inflammationis associated with ocular disease.

In one embodiment of the methods and uses of the present invention thedisease is a diabetes-induced disease selected from the group consistingof diabetic nephropathy, glomerulosclerosis, diabetic retinopathy,non-alcoholic fatty liver disease and choroidal neovascularisation.

In another embodiment of the methods and uses of the present inventionthe disease is a neuroinflammatory disease. In a further embodiment ofthe methods and uses of the present invention the disease is selectedfrom the group consisting of liver fibrosis, liver cirrhosis, kidneyfibrosis, idiopathic pulmonary fibrosis and radiation-induced fibrosis.In a still further embodiment of the methods and uses of the presentinvention the disease is cancer.

Definitions

The following are some definitions that may be helpful in understandingthe description of the present invention. These are intended as generaldefinitions and should in no way limit the scope of the presentinvention to those terms alone, but are put forth for a betterunderstanding of the following description.

Unless the context requires otherwise or specifically stated to thecontrary, integers, steps, or elements of the invention recited hereinas singular integers, steps or elements clearly encompass both singularand plural forms of the recited integers, steps or elements.

Throughout this specification, unless the context requires otherwise,the word “comprise”, or variations such as “comprises” or “comprising”,will be understood to imply the inclusion of a stated step or element orinteger or group of steps or elements or integers, but not the exclusionof any other step or element or integer or group of elements orintegers. Thus, in the context of this specification, the term“comprising” means “including principally, but not necessarily solely”.

Those skilled in the art will appreciate that the invention describedherein is susceptible to variations and modifications other than thosespecifically described. It is to be understood that the inventionincludes all such variations and modifications. The invention alsoincludes all of the steps, features, compositions and compounds referredto or indicated in this specification, individually or collectively, andany and all combinations or any two or more of said steps or features.

As used herein, the term “alkyl” includes within its meaning monovalent(“alkyl”) and divalent (“alkylene”) straight chain or branched chainsaturated hydrocarbon radicals having from 1 to 6 carbon atoms, e.g., 1,2, 3, 4, 5 or 6 carbon atoms (unless specifically defined). The straightchain or branched alkyl group is attached at any available point toproduce a stable compound. In many embodiments, a lower alkyl is astraight or branched alkyl group containing from 1-6, 1-4, or 1-2,carbon atoms. For example, the term alkyl includes, but is not limitedto, methyl, ethyl, 1-propyl, isopropyl, 1-butyl, 2-butyl, isobutyl,tert-butyl, amyl, 1,2-dimethylpropyl, 1,1-dimethylpropyl, pentyl,isopentyl, hexyl, 4-methylpentyl, 1-methylpentyl, 2-methylpentyl,3-methylpentyl, 2,2-dimethylbutyl, 3,3-dimethylbutyl, 1,2-dimethylbutyl,1,3-dimethylbutyl, 1,2,2-trimethylpropyl, 1,1,2-trimethylpropyl, and thelike.

The term “alkoxy” as used herein refers to straight chain or branchedalkyloxy (i.e, O-alkyl) groups, wherein alkyl is as defined above.Examples of alkoxy groups include methoxy, ethoxy, n-propoxy, andisopropoxy

The term “cycloalkyl” as used herein includes within its meaningmonovalent (“cycloalkyl”) and divalent (“cycloalkylene”) saturated,monocyclic, bicyclic, polycyclic or fused analogs. In the context of thepresent disclosure the cycloalkyl group may have from 3 to 10 or from 3to 7 carbon atoms A fused analog of a cycloalkyl means a monocyclic ringfused to an aryl or heteroaryl group in which the point of attachment ison the non-aromatic portion. Examples of cycloalkyl and fused analogsthereof include cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl,cycloheptyl, tetrahydronaphthyl, decahydronaphthyl, indanyl, and thelike.

The term “aryl” or variants such as “arylene” as used herein refers tomonovalent (“aryl”) and divalent (“arylene”) single, polynuclear,conjugated and fused analogs of aromatic hydrocarbons having from 6 to10 carbon atoms. A fused analog of aryl means an aryl group fused to amonocyclic cycloalkyl or monocyclic heterocyclyl group in which thepoint of attachment is on the aromatic portion. Examples of aryl andfused analogs thereof include phenyl, naphthyl, indanyl, indenyl,tetrahydronaphthyl, 2,3-dihydrobenzofuranyl, dihydrobenzopyranyl,1,4-benzodioxanyl, and the like. Examples of an arylene includephenylene and natpthylene. A “substituted aryl” is an aryl that isindependently substituted, with one or more, preferably 1, 2 or 3substituents, attached at any available atom to produce a stablecompound. A “substituted arylene” is an arylene that is independentlysubstituted, with one or more, preferably 1, 2 or 3 substituents,attached at any available atom to produce a stable compound.

The term “alkylaryl” as used herein, includes within its meaningmonovalent (“aryl”) and divalent (“arylene”), single, polynuclear,conjugated and fused aromatic hydrocarbon radicals attached to divalent,saturated, straight or branched chain alkylene radicals. Examples ofalkylaryl groups include, but are not limited to, benzyl.

The term “heteroaryl” refers to a monocyclic aromatic ring structurecontaining 5 or 6 ring atoms, wherein heteroaryl contains one or moreheteroatoms independently selected from the group consisting of O, S,and N. Heteroaryl is also intended to include oxidized S or N, such assulfinyl, sulfonyl and N-oxide of a tertiary ring nitrogen. A carbon ornitrogen atom is the point of attachment of the heteroaryl ringstructure such that a stable compound is produced. Examples ofheteroaryl groups include, but are not limited to, pyridinyl,pyridazinyl, pyrazinyl, quinaoxalyl, indolizinyl, benzo[b]thienyl,quinazolinyl, purinyl, indolyl, quinolinyl, pyrimidinyl, pyrrolyl,oxazolyl, thiazolyl, thienyl, isoxazolyl, oxathiadiazolyl, isothiazolyl,tetrazolyl, imidazolyl, triazinyl, furanyl, benzofuryl, and indolyl.“Nitrogen containing heteroaryl” refers to heteroaryl wherein anyheteroatoms are N. A “substituted heteroaryl” is a heteroaryl that isindependently substituted, with one or more, preferably 1, 2 or 3substituents, attached at any available atom to produce a stablecompound.

“Heteroarylene” refers to a divalent, monocyclic aromatic ring structurecontaining 5 or 6 ring atoms, wherein heteroarylene contains one or moreheteroatoms independently selected from the group consisting of O, S,and N. Heteroarylene is also intended to include oxidized S or N, suchas sulfinyl, sulfonyl and N-oxide of a tertiary ring nitrogen. A carbonor nitrogen atom is the point of attachment of the heteroarylene ringstructure to the substituents thereon, such that a stable compound isproduced. Examples of heteroaryl groups include, but are not limited to,pyridinylene, pyridazinylene, pyrazinylene, quinaoxalylene,indolizinylene, benzo[b]thienylene, quinazolinylene, purinylene,indolylene, quinolinylene, pyrimidinylene, pyrrolylene, oxazolylene,thiazolylene, thienylene, isoxazolylene, oxathiadiazolylene,isothiazolylene, tetrazolylene, imidazolylene, triazinylene, furanylene,benzofurylene, and indolylene. “Nitrogen containing heteroarylene”refers to heteroarylene wherein any heteroatoms are N. A “substitutedheteroarylene” is a heteroarylene that is independently substituted,with one or more, preferably 1, 2 or 3 substituents, attached at anyavailable atom to produce a stable compound.

The term “heterocyclyl” and variants such as “heterocycloalkyl” as usedherein, includes within its meaning monovalent (“heterocyclyl”) anddivalent (“heterocyclylene”), saturated, monocyclic, bicyclic,polycyclic or fused hydrocarbon radicals having from 3 to 10 ring atoms,wherein from 1 to 5, or from 1 to 3, ring atoms are heteroatomsindependently selected from O, N, NH, or S, in which the point ofattachment may be carbon or nitrogen. A fused analog of heterocyclylmeans a monocyclic heterocycle fused to an aryl or heteroaryl group inwhich the point of attachment is on the non-aromatic portion. Theheterocyclyl group may be C₃₋₈ heterocyclyl. The heterocycloalkyl groupmay be C₃₋₆ heterocyclyl. The heterocyclyl group may be C₃₋₅heterocyclyl. Examples of heterocyclyl groups and fused analogs thereofinclude aziridinyl, pyrrolidinyl, thiazolidinyl, piperidinyl,piperazinyl, imidazolidinyl, 2,3-dihydrofuro(2,3-b)pyridyl,benzoxazinyl, tetrahydroquinolinyl, tetrahydroisoquinolinyl,dihydroindolyl, quinuclidinyl, azetidinyl, morpholinyl,tetrahydrothiophenyl, tetrahydrofuranyl, tetrahydropyranyl, and thelike. The term also includes partially unsaturated monocyclic rings thatare not aromatic, such as 2- or 4-pyridones attached through thenitrogen or N-substituted uracils.

The term “halogen” or variants such as “halide” or “halo” as used hereinrefers to fluorine, chlorine, bromine and iodine.

The term “heteroatom” or variants such as “hetero-” or “heterogroup” asused herein refers to O, N, NH and S.

In general, “substituted” refers to an organic group as defined herein(e.g., an alkyl group) in which one or more bonds to a hydrogen atomcontained therein are replaced by a bond to non-hydrogen or non-carbonatoms. Substituted groups also include groups in which one or more bondsto a carbon(s) or hydrogen(s) atom are replaced by one or more bonds,including double or triple bonds, to a heteroatom. Thus, a substitutedgroup will be substituted with one or more substituents, unlessotherwise specified. In some embodiments, a substituted group issubstituted with 1, 2, 3, 4, 5, or 6 substituents.

The term “optionally substituted” as used herein means the group towhich this term refers may be unsubstituted, or may be substituted withone or more groups independently selected from alkyl, alkenyl, alkynyl,cycloalkyl, cycloalkenyl, heterocycloalkyl, halo, haloalkyl,haloalkynyl, hydroxyl, hydroxyalkyl, alkoxy, thioalkoxy, alkenyloxy,haloalkoxy, haloalkenyloxy, NO₂, NH(alkyl), N(alkyl)₂, nitroalkyl,nitroalkenyl, nitroalkynyl, nitroheterocyclyl, alkylamino, dialkylamino,alkenylamine, alkynylamino, acyl, alkenoyl, alkynoyl, acylamino,diacylamino, acyloxy, alkylsulfonyloxy, heterocycloxy, heterocycloamino,haloheterocycloalkyl, alkylsulfenyl, alkylcarbonyloxy, alkylthio,acylthio, phosphorus-containing groups such as phosphono and phosphinyl,aryl, heteroaryl, alkylaryl, aralkyl, alkylheteroaryl, cyano, cyanate,isocyanate, CO₂H, CO₂alkyl, C(O)NH₂, —C(O)NH(alkyl), and —C(O)N(alkyl)₂.Preferred substituents include halogen, C₁-C₆alkyl, C₂-C₆alkenyl,C₁-C₆haloalkyl, C₁-C₆alkoxy, hydroxy(C₁₋₆)alkyl, C₃-C₆cycloalkyl, C(O)H,C(O)OH, NHC(O)H, NHC(O)C₁-C₄alkyl, C(O)C₁-C₄alkyl, NH₂, NHC₁-C₄alkyl,N(C₁-C₄alkyl)₂, NO₂, OH and CN. Particularly preferred substituentsinclude C₁₋₃alkyl, C₁₋₃alkoxy, halogen, OH, hydroxy(C₁₋₃)alkyl (e.g.,CH₂OH), C(O)C₁-C₄alkyl (eg C(O)CH₃), and C₁₋₃haloalkyl (e.g, CF₃,CH₂CF₃).

The present invention includes within its scope all stereoisomeric andisomeric forms of the compounds disclosed herein, including alldiastereomeric isomers, racemates, enantiomers and mixtures thereof.Compounds of the present invention may have asymmetric centers and mayoccur, except when specifically noted, as mixtures of stereoisomers oras individual diastereomers, or enantiomers, with all isomeric formsbeing included in the present invention. It is also understood that thecompounds described by Formula I may be present as E and Z isomers, alsoknown as cis and trans isomers. Thus, the present disclosure should beunderstood to include, for example, E, Z, cis, trans, (R), (S), (L),(D), (+), and/or (−) forms of the compounds, as appropriate in eachcase. Where a structure has no specific stereoisomerism indicated, itshould be understood that any and all possible isomers are encompassed.Compounds of the present invention embrace all conformational isomers.Compounds of the present invention may also exist in one or moretautomeric forms, including both single tautomers and mixtures oftautomers. Also included in the scope of the present invention are allpolymorphs and crystal forms of the compounds disclosed herein.

The present invention includes within its scope isotopes of differentatoms. Any atom not specifically designated as a particular isotope ismeant to represent any stable isotope of that atom. Thus, the presentdisclosure should be understood to include deuterium and tritiumisotopes of hydrogen

All references cited in this application are specifically incorporatedby cross-reference in their entirety. Reference to any such documentsshould not be construed as an admission that the document forms part ofthe common general knowledge or is prior art.

In the context of this specification the term “administering” andvariations of that term including “administer” and “administration”,includes contacting, applying, delivering or providing a compound orcomposition of the invention to an organism, or a surface by anyappropriate means. In the context of this specification, the term“treatment”, refers to any and all uses which remedy a disease state orsymptoms, prevent the establishment of disease, or otherwise prevent,hinder, retard, or reverse the progression of disease or otherundesirable symptoms in any way whatsoever.

In the context of this specification the term “effective amount”includes within its meaning a sufficient but non-toxic amount of acompound or composition of the invention to provide a desired effect.Thus, the term “therapeutically effective amount” includes within itsmeaning a sufficient but non-toxic amount of a compound or compositionof the invention to provide the desired therapeutic effect. The exactamount required will vary from subject to subject depending on factorssuch as the species being treated, the sex, age and general condition ofthe subject, the severity of the condition being treated, the particularagent being administered, the mode of administration, and so forth.Thus, it is not possible to specify an exact “effective amount”.However, for any given case, an appropriate “effective amount” may bedetermined by one of ordinary skill in the art using only routineexperimentation.

BRIEF DESCRIPTION OF THE SEVERAL VIEWS OF THE DRAWINGS

FIGS. 1A-1E show the ability of Compound 23 to inhibit SSAO/VAP-1 enzymein various tissues in rats after a single oral dose, with activitydetermined 24 hours after administration: Abdominal Fat (FIG. 1A);Plasma (FIG. 1B); Lung (FIG. 1C); Aorta (FIG. 1D); Liver (FIG. 1E).

FIGS. 2A-2E show the ability of 2 mg/kg of Compound 23 to inhibitSSAO/VAP-1 enzyme in various tissues in rats after a single oral dose,with activity determined at various time points after administration:Abdominal Fat (FIG. 2A); Plasma (FIG. 2B); Lung (FIG. 2C); Aorta (FIG.2D); Liver (FIG. 2E).

FIGS. 3A-3E show the ability of Compound 23 to inhibit SSAO/VAP-1 enzymein various tissues in rats after 5 days of repeated, daily oral dosing,with activity determined 24 hours after administration of the finaldose: Abdominal Fat (FIG. 3A); Plasma (FIG. 3B); Lung (FIG. 3C); Aorta(FIG. 3D); Liver (FIG. 3E).

FIGS. 4A-4D show the ability of Compound 23 to reduce inflammation in aninflamed air pouch in a mouse model: Neutrophils in exudate (FIG. 4A);Exudate volume (FIG. 4B); IL-6 in exudate (FIG. 4C); TNF-α in exudate(FIG. 4D).

FIGS. 5A & 5B show the ability of Compound 23 to reduce leukocytemigration in the mouse cremaster microcirculation: Inhibition ofleukocyte rolling (FIG. 5A); Inhibition of leukocyte adhesion (FIG. 5B).

FIGS. 6A & 6B show the ability of Compound 23 to reduce inflammation ina cecal ligation and perforation (CLP) model in the mouse: Total cellcount (FIG. 6A); Reduction of mortality (FIG. 6B).

FIGS. 7A-7F show the ability of Compound 9 to reduce neutrophilmigration and microglial activation in a mouse model ofneurodegeneration: Neutrophils in substantia nigra (FIG. 7A);Neutrophils in dorso-lateral striatum (DLS) (FIG. 7B); Neutrophils inhippocampus (7C); Microglial cells in substantia nigra (FIG. 7D);Microglial cells in dorso-lateral striatum (DLS) (FIG. 7E); Microglialcells in hippocampus (FIG. 7F).

FIGS. 8A-8C show the ability of Compound 9 to reduce inflammation in amouse model of acute lung inflammation:

FIGS. 9A & 9B show the ability of Compound 23 to reduce neutrophilmigration to the lung and airway hyper reactivity in a mouse model ofallergic asthma: Inhibition of neutrophils in broncheoalveolar lavagefluid (BALF)(FIG. 9A); Inhibition of methacholine-induced hyperreactivity (FIG. 9B).

FIGS. 10A & 10B show the ability of Compound 9 to reduce leukocytemigration into the lung and protect against mortality in a mouse modelof bacterial lung infection: Reduction of mortality (FIG. 10A);Inhibition of leukocytes in broncheoalveloar lavage fluid (BALF) (FIG.10B).

FIG. 11 shows the ability of Compound 23 to reduce the amount of solublecollagen in a mouse model of COPD.

FIGS. 12A-12E show the ability of Compound 23 to improve liver function,reduce fibrosis and reduce inflammation in a rat model of liverfibrosis; Reduction in serum ALT (FIG. 12A); Reduction in liver AST(FIG. 12B); Reduction in Sirius red area (FIG. 12C); Reduction ofinflammation score (FIG. 12D); Reduction of steatotic regions in theliver (FIG. 12E).

FIGS. 13A-13D show the ability of Compound 23 to reduce inflammation andfibrosis in a mouse model of fatty liver disease: Reduction ofinflammation score (FIG. 13A); Reduction in non-alcoholic fatty liverdisease (NAFLD) activity score (FIG. 13B); Reduction in Sirius red area(FIG. 13C); Reduction in liver triglycerides (FIG. 13D).

FIGS. 14A & 14B show the ability of Compound 23 to reduce inflammationin a mouse model of uveitis: Reduction of clinical score (FIG. 14A);Reduction of eosinophil infiltration (FIG. 14B).

DETAILED DESCRIPTION

The present invention relates to substituted haloallylamine compoundsthat may inhibit SSAO/VAP-1.

In accordance with the present invention, there are provided compoundshaving the structure (Formula I):

or a stereoisomer, pharmaceutically acceptable salt, polymorphic form,solvate or prodrug thereof; wherein:R¹ and R⁴ are independently hydrogen or optionally substitutedC₁₋₆alkyl;R² and R³ are independently selected from the group consisting ofhydrogen, chlorine and fluorine; provided, however, that R² and R³ arenot hydrogen at the same time;R⁵ is an optionally substituted arylene group;R⁶ is selected from

R⁷ and R⁸ are independently selected from the group consisting ofhydrogen, optionally substituted C₁₋₆alkyl and optionally substitutedC₃₋₇cycloalkyl; andX is CH₂, oxygen, sulfur or SO₂.

In one embodiment of compounds of the present invention R¹ and R⁴ areboth hydrogen. In another embodiment of compounds of the presentinvention R¹ is hydrogen and R⁴ is optionally substituted C₁₋₆alkyl. Ina further embodiment of compounds of the present invention R¹ isoptionally substituted C₁₋₆alkyl and R⁴ is hydrogen. In anotherembodiment of compounds of the present invention R¹ is hydrogen and R⁴is methyl. In a further embodiment of compounds of the present inventionR¹ is methyl and R⁴ is hydrogen.

In one embodiment of compounds of the present invention R² and R³ areindependently selected from the group consisting of hydrogen, chlorineand fluorine, provided that R² and R³ are not hydrogen at the same time.In another embodiment of compounds of the present invention R² and R³are independently hydrogen or fluorine, provided that R² and R³ are nothydrogen at the same time. In a further embodiment of compounds of thepresent invention R² and R³ are both fluorine. In another embodiment ofcompounds of the present invention R² is hydrogen and R³ is fluorine. Ina further embodiment of compounds of the present invention R² isfluorine and R³ is hydrogen.

In one embodiment of compounds of the present invention R⁵ is anoptionally substituted arylene group. In another embodiment of compoundsof the present invention R⁵ is an unsubstituted arylene group. In afurther embodiment of compounds of the present invention R⁵ is anoptionally substituted phenylene group. In another embodiment ofcompounds of the present invention R⁵ is an unsubstituted phenylenegroup. In one embodiment of compounds of the present invention R⁵ is aphenylene group optionally substituted by one or more groupsindependently selected from alkyl, halo, alkoxy and haloalkyl. Inanother embodiment of compounds of the present invention R⁵ is aphenylene group optionally substituted by one or more groupsindependently selected from methyl, fluorine, chlorine, bromine, OCH₃and CF₃.

In one embodiment of compounds of the present invention R⁶ is selectedfrom:

In another embodiment of compounds of the present invention R⁶ is

In a further embodiment of compounds of the present invention R⁶ is

In one embodiment of compounds of the present invention R⁷ and R⁸ areindependently selected from the group consisting of hydrogen, optionallysubstituted C₁₋₆alkyl and optionally substituted C₃₋₇cycloalkyl. Inanother embodiment of compounds of the present invention R⁷ and R⁸ areindependently selected from the group consisting of hydrogen andoptionally substituted C₁₋₆alkyl. In a further embodiment of compoundsof the present invention R⁷ and R⁸ are both hydrogen. In anotherembodiment of compounds of the present invention R⁷ and R⁸ are bothC₁₋₆alkyl. In a further embodiment of compounds of the present inventionR⁷ is hydrogen and R⁸ is C₁₋₆alkyl. In a still further embodiment R⁷ andR⁸ are independently selected from the group consisting of hydrogen,tert-butyl, methyl, ethyl, isopropyl and 2-butyl.

In one embodiment of compounds of the present invention X is CH₂,oxygen, sulfur or SO₂. In another embodiment of compounds of the presentinvention X is CH₂, oxygen or sulfur. In further embodiment of compoundsof the present invention X is oxygen.

In a particular embodiment of the present invention, there is provided acompound having the structure (Formula II), as follows:

or a pharmaceutically acceptable salt, solvate, polymorphic form, orprodrug thereof; wherein:R⁵ is an optionally substituted arylene group;R⁶ is selected from

R⁷ and R⁸ are independently selected from the group consisting ofhydrogen, optionally substituted C₁₋₆alkyl and optionally substitutedC₃₋₇cycloalkyl; andX is CH₂, oxygen, sulfur or SO₂.

In accordance with one embodiment of the present invention, presentlypreferred compounds include compounds of Formulae I and II wherein R³ isfluorine, and X is oxygen.

It is understood that compounds described by Formulae I or II may beadministered in a prodrug form wherein the substituent R¹ can beselected from such functional groups as —C(O)alkyl, —C(O)aryl,—C(O)-arylalkyl, C(O)heteroaryl, —C(O)— heteroarylalkyl, or the like.

The compounds described by Formula I may exist as acid addition saltswhen a basic amino group is present, or as metal salts when an acidicgroup is present.

Exemplary compounds according to the present invention include thecompounds set forth in Table 1:

TABLE 1  1

(Z)-4-(2-(Aminomethyl)-3- fluoroallyloxy)-N-tert- butylbenzamide  2

(Z)-4-(2-(Aminomethyl)-3- fluoroallyloxy)benzamide  3

(E)-4-(2-(Aminomethyl)-3- fluoroallyloxy)benzamide  4

(Z)-4-(2-(Aminomethyl)-3- fluoroallyloxy)-3-fluoro-N,N-dimethylbenzamide  5

(E)-4-(3-(Aminomethyl)-4- fluorobut-3-en-2-yloxy)-N-tert- butylbenzamide 6

(Z)-4-(2-(Aminomethyl)-3- fluoroallyloxy)-3-chloro-N,N-dimethylbenzamide  7

4-(2-(Aminomethyl)-3- fluoroallyloxy)-3-methoxy-N,N- dimethylbenzamide 8

4-(2-(Aminomethyl)-3- fluoroallylthio)-N,N- dimethylbenzamide  9

(Z)-4-(2-(Aminomethyl)-3- fluoroallyloxy)benzene- sulfonamide 10

(Z)-4-(2-(Aminomethyl)-3- fluoroallyloxy)-N,N-dimethylbenzenesulfonamide 11

(E)-4-(2-(Aminomethyl)-3- fluoroallyloxy)benzene- sulfonamide 12

(E)-N-tert-Butyl-4-(3-fluoro-2- ((methylamino)methyl)allyloxy) benzamide13

(E)-4-(2-(Aminomethyl)-3- fluoroallyloxy)-N,N- dimethylbenzamide 14

(E)-4-(2-(Aminomethyl)-3- fluoroallyloxy)-N,N-dimethylbenzenesulfonamide 15

(Z)-3-(2-(Aminomethyl)-3- fluoroallyloxy)-N,N-dimethylbenzenesulfonamide 16

(Z)-4-(2-(Aminomethyl)-3- fluoroallyloxy)-N-tert-butylbenzenesulfonamide 17

(E)-4-(2-(Aminomethyl)-3- fluoroallyloxy)-N-tert-butylbenzenesulfonamide 18

(Z)-4-(2-(Aminomethyl)-3- fluoroallyloxy)-N,N- dimethylbenzamide 19

(E)-4-(2-(Aminomethyl)-3- fluoroallyloxy)-N-tert-butyl-3-fluorobenzamide 20

(Z)-4-(2-(Aminomethyl)-3- fluoroallyloxy)-3-bromo-N,N- dimethylbenzamide21

(E)-4-(2-(Aminomethyl)-3- fluoroallyloxy)-N-tert-butyl-2-(trifluoromethyl)benzamide 22

(E)-4-(2-(Aminomethyl)-3- chloroallyloxy)-N-tert- butylbenzamide 23

(E)-4-(2-(Aminomethyl)-3- fluoroallyloxy)-N-tert- butylbenzamide 24

(E)-4-(2-(Aminomethyl)-3- fluoroallyloxy)-N,N- diethylbenzamide 25

(E)-4-(2-(Aminomethyl)-3- fluoroallyloxy)-N- methylbenzamide 26

(Z)-4-(2-(Aminomethyl)-3- fluoroallyloxy)-N,N,2- trimethylbenzamide 27

(Z)-4-(2-(Aminomethyl)-3- chloroallyloxy)-N-tert- butylbenzamide 28

(E)-4-(2-(Aminomethyl)-3- fluoroallyloxy)-N- methylbenzenesulfonamide 29

(Z)-4-(2-(Aminomethyl)-3- fluoroallyloxy)-N- methylbenzenesulfonamide 30

(E)-4-(2-(Aminomethyl)-3- fluoroallyloxy)-N- ethylbenzenesulfonamide 31

(Z)-4-(2-(Aminomethyl)-3- fluoroallyloxy)-N- ethylbenzenesulfonamide 32

(E)-4-(2-(Aminomethyl)-3- fluoroallyloxy)-N- isopropylbenzenesulfonamide33

(Z)-4-(2-(Aminomethyl)-3- fluoroallyloxy)-N- isopropylbenzenesulfonamide34

(Z)-4-(3-(Aminomethyl)-4- fluorobut-3-enyl)-N-tert- butylbenzamide 35

(Z)-4-(2-(Aminomethyl)-3- fluoroallyloxy)-N-ethyl-N- methylbenzamide 36

(Z)-4-(2-(Aminomethyl)-3- fluoroallyloxy)-N-sec-butyl-N- methylbenzamide37

(Z)-4-(2-(Aminomethyl)-3- fluoroallyloxy)-N-tert-butyl-N-methylbenzenesulfonamide 38

(Z)-4-(2-(Aminomethyl)-3- fluoroallyloxy)-N-isopropyl-N-methylbenzenesulfonamide 39

(E)-4-(2-(Aminomethyl)-3- fluoroallyloxy)-N- isopropylbenzamideor a pharmaceutically acceptable salt or solvate thereof.Preparation of Compounds of Formula I

The compounds of the invention can be prepared in a variety of ways,such as, for example, procedures described in U.S. Pat. No. 4,454,158;U.S. Pat. No. 4,699,928; and U.S. Pat. No. 4,650,907.

An alternate route to prepare compounds described by Formula I in whichX═O or S employs the synthetic protocol described in Scheme 1, below.This is similar to procedures described in WO 2007/120528.

wherein R², R³, X and R⁵ are as defined herein; P₁ is a functional groupused to protect a nitrogen functionality; and LG is a leaving group.Examples of P₁ are carbonates such as the tert-butyloxycarbonyl (BOC),the 9-fluorenylmethyloxy-carbonyl (FMOC), and the benzyloxycarbonyl(CBZ) groups; examples of LG are bromo, chloro, iodo, triflates,tosylates, mesylates, and ester groups.

A compound represented by Formula III is either directly used in adisplacement reaction (Method A), such as a Mitsunobu reaction, to yieldthe compound represented by Formula IV, or is first converted to acompound represented by Formula V which contains a leaving group (LG),such as bromide, chloride or iodide, by procedures well known in the art(Method B). Alternatively that alcohol can be directly activated withthe tosyl protecting/activating group (P₂=Tosyl in Scheme 2, FormulaVIII; see below). The activated compound described by Formula V is thentreated with a nucleophilic reagent to furnish the compound representedby Formula IV (Method C).

The Mitsunobu reaction conditions are well described in the scientificand patent literature (available on the world wide web aten.wikipedia.org/wiki/Mitsunobu_reaction, and Mitsunobu, O. The use ofdiethyl azodicarboxylate and triphenylphosphine in synthesis andtransformation of natural products. Synthesis 1981, 1-28) and proceed bycontacting an alcohol with an appropriately substituted phenolic orthiophenolic group, or a substituted phthalimide in the presence of adialkyl azodicarboxylate and triphenylphosphine in an organic solventsuch as tetrahydrofuran (THF) or CH2Cl2 (CH₂Cl₂).

Conversion of the alcohol group in Formula III to the correspondingbromide, chloride or iodide is accomplished by any number of commonlyused procedures (See, for example, March J. Advanced Organic Synthesis,John Wiley & Sons, Third Edition 1985), including treatment with PBr₃ intoluene or CBr₄ and triphenylphosphine in an organic solvent such asCH₂Cl₂. The resulting halide can be treated with nucleophiles such assubstituted alcohols, phenols, amines, or thiols to afford the compoundrepresented by Formula IV.

There are many well established chemical procedures for the deprotectionof the compounds described by Formula IV to the inventive compoundsdescribed by Formula I (Method J; see Scheme 2). For example if P₁ is aBOC protecting group, compounds described by Formula IV can be treatedwith an acidic substance such as dry hydrogen chloride in a solvent suchas diethyl ether to furnish the compounds described by Formula I as thehydrochloride salt. In general, the free amino compounds are convertedto acid addition salts for ease of handling and for improved chemicalstability. Examples of acid addition salts include but are not limitedto hydrochloride, hydrobromide and methanesulfonate salts.

The preparation of compounds described by Formula III is straightforwardfrom either commercially available or readily accessible aminodiolillustrated by Formula VI (See Scheme 3).

The first step is selective protection of the primary amine, preferablyas the tert-butyl carbamate (BOC) (P₁═BOC in Formula VII), followed byselective protection of the primary alcohol to afford the alcoholdescribed by Formula IX. Selective protection methods (Method E) arewell known in the art of synthetic chemistry. For example, the primaryalcohol can be selectively reacted withtert-butyl-(chloro)dimethylsilane in the presence of imizadole tofurnish the tert-butyldimethylsilyl protected alcohol (Formula VII).Oxidation of the secondary alcohol is best achieved under Swernoxidation conditions (Method F) resulting in the ketone represented byFormula VIII. The haloalkene functional group in Formula X is introducedby Wittig or Horner-Wadsworth-Emmons reaction. When R² and R³ are F andH in the structure described by Formula I, reaction of the ketonedescribed by Formula VIII with fluoromethyl (triphenyl)phosphoniumtetrafluoroborate in the presence of a strong base such as sodiumbis(trimethylsilyl) amide affords the fluoroalkene as a mixture of E andZ isomers (described by Formula X). These isomers can be separated bychromatographic procedures to afford the individual E and Z isomers.Removal of the protecting group in the compounds described by Formula Xcan be readily achieved (Method H). The choice of the deprotectingreagent is determined by the nature of the protecting groups P₁ and P₂.When P₂ is tert-butyldimethylsilyl and P₁ is the BOC group, selectiveremoval of P₂ is achieved with TBAF to yield the alcohol described byFormula III.

Therapeutic Uses and Formulations

The present invention provides methods for the use of compoundsdescribed by Formulae I and II to inhibit membrane-bound SSAO/VAP-1 andsoluble SSAO/VAP-1. The relative inhibitory potencies of the compoundscan be determined by the amount needed to inhibit the amine oxidaseactivity of SSAO/VAP-1 in a variety of ways, e.g., in an in vitro assaywith recombinant human protein or with recombinant non-human enzyme, incellular assays expressing normal rodent enzyme, in cellular assayswhich have been transfected with human protein, in in vivo tests inrodent and other mammalian species, and the like.

The present invention also discloses methods to use the compoundsdescribed by Formulae I and II to inhibit SSAO/VAP-1 in patientssuffering from an inflammatory disease, and methods to treatinflammatory diseases. Human inflammatory diseases include arthritis,Crohn's disease, irritable bowel disease, psoriasis, asthma, chronicpulmonary obstructive disease, bronchiectasis, arthrosclerosis,inflammation due to diabetes, and inflammatory cell destructionfollowing stroke.

Thus, in one aspect, the present invention is directed to methods ofinhibiting an amine oxidase enzyme in a subject in need thereof, saidmethods comprising administering to said subject an effective amount ofa compound of Formula I or Formula II to effect a positive therapeuticresponse.

In another aspect, the present invention is directed to methods oftreating a disease associated with an amine oxidase enzyme, said methodscomprising administering to a subject in need thereof a therapeuticallyeffective amount of a compound of Formula I or Formula II.

In still another aspect, the present invention is directed to methods oftreating a disease modulated by SSAO/VAP-1, said methods comprisingadministering to a subject in need thereof a therapeutically effectiveamount of a compound of Formula I or Formula II.

The above-described methods are applicable wherein the disease isinflammation. As employed herein, “inflammation” embraces a wide varietyof indications, including arthritis (including juvenile rheumatoidarthritis), Crohn's disease, ulcerative colitis, inflammatory boweldiseases (e.g., irritable bowel disease), psoriasis, asthma, pulmonaryinflammation, chronic pulmonary obstructive disease (COPD),bronchiectasis, skin inflammation, ocular disease, contact dermatitis,liver inflammation, liver autoimmune diseases, autoimmune hepatitis,primary biliary cirrhosis, sclerosing cholangitis, autoimmunecholangitis, alcoholic liver disease, artherosclerosis, chronic heartfailure, congestive heart failure, ischemic diseases, stroke andcomplications thereof, myocardial infarction and complications thereof,inflammatory cell destruction following stroke, synovitis, systemicinflammatory sepsis, and the like.

The above-described methods are also applicable wherein the disease isType I diabetes and complications thereof, Type II diabetes andcomplications thereof, and the like.

The above described methods are also applicable wherein the disease ismacular degeneration or other ocular diseases.

The above described methods are also applicable wherein the disease isfibrosis. As employed here “fibrosis” includes such diseases as cysticfibrosis, idiopathic pulmonary fibrosis, liver fibrosis, includingnon-alcoholic fatty liver diseases such as non-alcoholic steatohepatitis(NASH) and alcohol induced fibrosis leading to cirrhosis of the liver,kidney fibrosis, scleroderma, radiation-induced fibrosis and otherdiseases where excessive fibrosis contributes to disease pathology.

The above-described methods are also applicable wherein the disease is aneuroinflammatory disease. As employed herein, “neuroinflammatorydiseases” embrace a variety of indications, including stroke,Parkinson's disease, Alzheimer's disease, vascular dementia, multiplesclerosis, chronic multiple sclerosis, and the like.

The above-described methods are also applicable wherein the disease iscancer. In one embodiment the cancer is selected from the groupconsisting of lung cancer; breast cancer; colorectal cancer; analcancer; pancreatic cancer; prostate cancer; ovarian carcinoma; liver andbile duct carcinoma; esophageal carcinoma; non-Hodgkin's lymphoma;bladder carcinoma; carcinoma of the uterus; glioma, glioblastoma,medullablastoma, and other tumors of the brain; kidney cancer; cancer ofthe head and neck; cancer of the stomach; multiple myeloma; testicularcancer; germ cell tumor; neuroendocrine tumor; cervical cancer;carcinoids of the gastrointestinal tract, breast, and other organs;signet ring cell carcinoma; mesenchymal tumors including sarcomas,fibrosarcomas, haemangioma, angiomatosis, haemangiopericytoma,pseudoangiomatous stromal hyperplasia, myofibroblastoma, fibromatosis,inflammatory myofibroblastic tumour, lipoma, angiolipoma, granular celltumour, neurofibroma, schwannoma, angiosarcoma, liposarcoma,rhabdomyosarcoma, osteosarcoma, leiomyoma or a leiomysarcoma.

Pharmaceutical and/or Therapeutic Formulations

In another embodiment of the present invention, there are providedcompositions comprising a compound having Formula I or Formula II and atleast one pharmaceutically acceptable excipient, carrier or diluenttherefor. The compounds of Formula I may also be present as suitablesalts, including pharmaceutically acceptable salts.

The phrase “pharmaceutically acceptable carrier” refers to any carrierknown to those skilled in the art to be suitable for the particular modeof administration. In addition, the compounds may be formulated as thesole pharmaceutically active ingredient in the composition or may becombined with other active ingredients.

The phrase “pharmaceutically acceptable salt” refers to any saltpreparation that is appropriate for use in a pharmaceutical application.By pharmaceutically acceptable salt it is meant those salts which,within the scope of sound medical judgement, are suitable for use incontact with the tissues of humans and lower animals without unduetoxicity, irritation, allergic response and the like, and arecommensurate with a reasonable benefit/risk ratio. Pharmaceuticallyacceptable salts are well known in the art and include acid addition andbase salts. Hemisalts of acids and bases may also be formed.Pharmaceutically-acceptable salts include amine salts of mineral acids(e.g., hydrochlorides, hydrobromides, sulfates, and the like); and aminesalts of organic acids (e.g., formates, acetates, lactates, malates,tartrates, citrates, ascorbates, succinates, maleates, butyrates,valerates, fumarates, and the like).

For compounds of formula (I) having a basic site, suitablepharmaceutically acceptable salts may be acid addition salts. Forexample, suitable pharmaceutically acceptable salts of such compoundsmay be prepared by mixing a pharmaceutically acceptable acid such ashydrochloric acid, sulfuric acid, methanesulfonic acid, succinic acid,fumaric acid, maleic acid, benzoic acid, phosphoric acid, acetic acid,oxalic acid, carbonic acid, tartaric acid, or citric acid with thecompounds of the invention.

S. M. Berge et al. describe pharmaceutically acceptable salts in detailin J. Pharmaceutical Sciences, 1977, 66:1-19. The salts can be preparedin situ during the final isolation and purification of the compounds ofthe invention, or separately by reacting the free base function with asuitable organic acid. Representative acid addition salts includeacetate, adipate, alginate, ascorbate, asparate, benzenesulfonate,benzoate, bisulfate, borate, butyrate, camphorate, camphorsulfonate,citrate, digluconate, cyclopentanepropionate, dodecylsulfate,ethanesulfonate, fumarate, glucoheptonate, glycerophosphate,hemisulfate, heptonate, hexanoate, hydrobromide, hydrochloride,hydroiodide, 2-hydroxy-ethanesulfonate, lactobionate, lactate, laurate,lauryl sulfate, malate, maleate, malonate, methanesulfonate,2-naphthalenesulfonate, nicotinate, nitrate, oleate, oxalate, palmitate,pamoate, pectinate, persulfate, 3-phenylpropionate, phosphate, picrate,pivalate, propionate, stearate, succinate, sulfate, tartrate,thiocyanate, toluenesulfonate, undecanoate, valerate salts, and thelike. Suitable base salts are formed from bases that form non-toxicsalts. Examples include the aluminium, arginine, benzathine, calcium,choline, diethylamine, diolamine, glycine, lysine, magnesium, meglumine,olamine, potassium, sodium, tromethamine and zinc salts. Representativealkali or alkaline earth metal salts include sodium, lithium potassium,calcium, magnesium, and the like, as well as non-toxic ammonium,quaternary ammonium, and amine cations, including, but not limited toammonium, tetramethylammonium, tetraethylammonium, methylamine,dimethylamine, trimethylamine, triethylamine, ethylamine,triethanolamine and the like.

Pharmaceutically acceptable salts of compounds of formula I may beprepared by methods known to those skilled in the art, including forexample

-   -   i. by reacting the compound of formula I with the desired acid        or base;    -   ii. by removing an acid- or base-labile protecting group from a        suitable precursor of the compound of formula I or by        ring-opening a suitable cyclic precursor, for example, a lactone        or lactam, using the desired acid or base; or    -   iii. by converting one salt of the compound of formula I to        another by reaction with an appropriate acid or base or by means        of a suitable ion exchange column.

The above reactions (i)-(iii) are typically carried out in solution. Theresulting salt may precipitate out and be collected by filtration or maybe recovered by evaporation of the solvent. The degree of ionisation inthe resulting salt may vary from completely ionised to almostnon-ionised.

Thus, for instance, suitable pharmaceutically acceptable salts ofcompounds according to the present invention may be prepared by mixing apharmaceutically acceptable acid such as hydrochloric acid, sulfuricacid, methanesulfonic acid, succinic acid, fumaric acid, maleic acid,benzoic acid, phosphoric acid, acetic acid, oxalic acid, carbonic acid,tartaric acid, or citric acid with the compounds of the invention.Suitable pharmaceutically acceptable salts of the compounds of thepresent invention therefore include acid addition salts.

The compounds of the invention may exist in both unsolvated and solvatedforms. The term ‘solvate’ is used herein to describe a molecular complexcomprising the compound of the invention and a stoichiometric amount ofone or more pharmaceutically acceptable solvent molecules, for example,ethanol. The term ‘hydrate’ is employed when the solvent is water.

In one embodiment the compounds of Formula I may be administered in theform of a “prodrug”. The phrase “prodrug” refers to a compound that,upon in vivo administration, is metabolized by one or more steps orprocesses or otherwise converted to the biologically, pharmaceuticallyor therapeutically active form of the compound. Prodrugs can be preparedby modifying functional groups present in the compound in such a waythat the modifications are cleaved, either in routine manipulation or invivo, to a compound described herein. For example, prodrugs includecompounds of the present invention wherein a hydroxy, amino, orsulfhydryl group is bonded to any group that, when administered to amammalian subject, can be cleaved to form a free hydroxyl, free amino,or free sulfhydryl group, respectively. Representative prodrugs include,for example, amides, esters, enol ethers, enol esters, acetates,formates, benzoate derivatives, and the like of alcohol and aminefunctional groups in the compounds of the present invention. By virtueof knowledge of pharmacodynamic processes and drug metabolism in vivo,those of skill in this art, once a pharmaceutically active compound isknown, can design prodrugs of the compound (see, e.g., Nogrady (1985)Medicinal Chemistry A Biochemical Approach, Oxford University Press, NewYork, pages 388-392).

Compositions herein comprise one or more compounds provided herein. Thecompounds are, in one embodiment, formulated into suitablepharmaceutical preparations such as solutions, suspensions, tablets,dispersible tablets, pills, capsules, powders, sustained releaseformulations or elixirs, for oral administration or in sterile solutionsor suspensions for parenteral administration, as well as transdermalpatch preparation and dry powder inhalers. In one embodiment, thecompounds described above are formulated into pharmaceuticalcompositions using techniques and procedures well known in the art (see,e.g., Ansel Introduction to Pharmaceutical Dosage Forms, Fourth Edition1985, 126).

In the compositions, effective concentrations of one or more compoundsor pharmaceutically acceptable derivatives thereof is (are) mixed with asuitable pharmaceutical carrier. The compounds may be derivatized as thecorresponding salts, esters, enol ethers or esters, acetals, ketals,orthoesters, hemiacetals, hemiketals, acids, bases, solvates, hydratesor prodrugs prior to formulation, as described above. The concentrationsof the compounds in the compositions are effective for delivery of anamount, upon administration, that treats, prevents, or ameliorates oneor more of the symptoms of diseases or disorders to be treated.

In one embodiment, the compositions are formulated for single dosageadministration. To formulate a composition, the weight fraction ofcompound is dissolved, suspended, dispersed or otherwise mixed in aselected carrier at an effective concentration such that the treatedcondition is relieved, prevented, or one or more symptoms areameliorated.

The active compound is included in the pharmaceutically acceptablecarrier in an amount sufficient to exert a therapeutically useful effectin the absence of undesirable side effects on the patient treated. Thetherapeutically effective concentration may be determined empirically bytesting the compounds in in vitro and in vivo systems described hereinand in PCT publication WO 04/018997, and then extrapolated therefrom fordosages for humans.

The concentration of active compound in the pharmaceutical compositionwill depend on absorption, inactivation and excretion rates of theactive compound, the physicochemical characteristics of the compound,the dosage schedule, and amount administered as well as other factorsknown to those of skill in the art.

In one embodiment, a therapeutically effective dosage should produce aserum concentration of active ingredient of from about 0.1 ng/mL toabout 50-100 μg/mL. The pharmaceutical compositions, in anotherembodiment, should provide a dosage of from about 0.001 mg to about 2000mg of compound per kilogram of body weight per day. Pharmaceuticaldosage unit forms are prepared to provide from about 0.01 mg, 0.1 mg or1 mg to about 500 mg, 1000 mg or 2000 mg, and in one embodiment fromabout 10 mg to about 500 mg of the active ingredient or a combination ofessential ingredients per dosage unit form.

Dosing may occur at intervals of minutes, hours, days, weeks, months oryears or continuously over any one of these periods. Suitable dosageslie within the range of about 0.1 ng per kg of body weight to 1 g per kgof body weight per dosage. The dosage is preferably in the range of 1 μgto 1 g per kg of body weight per dosage, such as is in the range of 1 mgto 1 g per kg of body weight per dosage. Suitably, the dosage is in therange of 1 μg to 500 μg per kg of body weight per dosage, such as 1 μgto 200 mg per kg of body weight per dosage, or 1 μg to 100 mg per kg ofbody weight per dosage. Other suitable dosages may be in the range of 1mg to 250 mg per kg of body weight, including 1 mg to 10, 20, 50 or 100mg per kg of body weight per dosage or 10 μg to 100 mg per kg of bodyweight per dosage.

Suitable dosage amounts and dosing regimens can be determined by theattending physician and may depend on the particular condition beingtreated, the severity of the condition, as well as the general health,age and weight of the subject.

The active ingredient may be administered at once, or may be dividedinto a number of smaller doses to be administered at intervals of time.It is understood that the precise dosage and duration of treatment is afunction of the disease being treated and may be determined empiricallyusing known testing protocols or by extrapolation from in vivo or invitro test data. It is to be noted that concentrations and dosage valuesmay also vary with the severity of the condition to be alleviated. It isto be further understood that for any particular subject, specificdosage regimens should be adjusted over time according to the individualneed and the professional judgment of the person administering orsupervising the administration of the compositions, and that theconcentration ranges set forth herein are exemplary only and are notintended to limit the scope or practice of the claimed compositions.

In instances in which the compounds exhibit insufficient solubility,methods for solubilizing compounds may be used. Such methods are knownto those of skill in this art, and include, but are not limited to,using co-solvents, such as dimethylsulfoxide (DMSO), using surfactants,such as TWEEN®, dissolution in aqueous sodium bicarbonate, formulatingthe compounds of interest as nanoparticles, and the like. Derivatives ofthe compounds, such as prodrugs of the compounds may also be used informulating effective pharmaceutical compositions.

Upon mixing or addition of the compound(s), the resulting mixture may bea solution, suspension, emulsion or the like. The form of the resultingmixture depends upon a number of factors, including the intended mode ofadministration and the solubility of the compound in the selectedcarrier or vehicle. The effective concentration is sufficient forameliorating the symptoms of the disease, disorder or condition treatedand may be empirically determined.

The pharmaceutical compositions are provided for administration tohumans and animals in unit dosage forms, such as tablets, capsules,pills, powders, granules, sterile parenteral solutions or suspensions,and oral solutions or suspensions, and oil-water emulsions containingsuitable quantities of the compounds or pharmaceutically acceptablederivatives thereof. The pharmaceutically therapeutically activecompounds and derivatives thereof are, in one embodiment, formulated andadministered in unit-dosage forms or multiple-dosage forms. Unit-doseforms as used herein refers to physically discrete units suitable forhuman and animal subjects and packaged individually as is known in theart. Each unit-dose contains a predetermined quantity of thetherapeutically active compound sufficient to produce the desiredtherapeutic effect, in association with the required pharmaceuticalcarrier, vehicle or diluent. Examples of unit-dose forms includeampoules and syringes and individually packaged tablets or capsules.Unit-dose forms may be administered in fractions or multiples thereof. Amultiple-dose form is a plurality of identical unit-dosage formspackaged in a single container to be administered in segregatedunit-dose form. Examples of multiple-dose forms include vials, bottlesof tablets or capsules or bottles of pints or gallons. Hence, multipledose form is a multiple of unit-doses which are not segregated inpackaging.

Actual methods of preparing such dosage forms are known, or will beapparent, to those skilled in this art; for example, see Remington'sPharmaceutical Sciences, Mack Publishing Company, Easton, Pa., 15thEdition, 1975.

Dosage forms or compositions containing active ingredient in the rangeof 0.005% to 100% (wt %) with the balance made up from non-toxic carriermay be prepared. Methods for preparation of these compositions are knownto those skilled in the art. The contemplated compositions may contain0.001%-100% (wt %) active ingredient, in one embodiment 0.1-95% (wt %),in another embodiment 75-85% (wt %).

Modes of Administration

Convenient modes of administration include injection (subcutaneous,intravenous, etc.), oral administration, inhalation, transdermalapplication, topical creams or gels or powders, vaginal or rectaladministration. Depending on the route of administration, theformulation and/or compound may be coated with a material to protect thecompound from the action of enzymes, acids and other natural conditionswhich may inactivate the therapeutic activity of the compound. Thecompound may also be administered parenterally or intraperitoneally.

Compositions for Oral Administration

Oral pharmaceutical dosage forms are either solid, gel or liquid. Thesolid dosage forms are tablets, capsules, granules, and bulk powders.Types of oral tablets include compressed, chewable lozenges and tabletswhich may be enteric-coated, sugar-coated or film-coated. Capsules maybe hard or soft gelatin capsules, while granules and powders may beprovided in non-effervescent or effervescent form with the combinationof other ingredients known to those skilled in the art.

Solid Compositions for Oral Administration

In certain embodiments, the formulations are solid dosage forms, in oneembodiment, capsules or tablets. The tablets, pills, capsules, trochesand the like can contain one or more of the following ingredients, orcompounds of a similar nature: a binder; a lubricant; a diluent; aglidant; a disintegrating agent; a colouring agent; a sweetening agent;a flavouring agent; a wetting agent; an emetic coating; and a filmcoating. Examples of binders include microcrystalline cellulose, gumtragacanth, glucose solution, acacia mucilage, gelatin solution,molasses, polvinylpyrrolidine, povidone, crospovidones, sucrose andstarch paste. Lubricants include talc, starch, magnesium or calciumstearate, lycopodium and stearic acid. Diluents include, for example,lactose, sucrose, starch, kaolin, salt, mannitol and dicalciumphosphate. Glidants include, but are not limited to, colloidal silicondioxide. Disintegrating agents include crosscarmellose sodium, sodiumstarch glycolate, alginic acid, corn starch, potato starch, bentonite,methylcellulose, agar and carboxymethylcellulose. Coloring agentsinclude, for example, any of the approved certified water soluble FD andC dyes, mixtures thereof; and water insoluble FD and C dyes suspended onalumina hydrate. Sweetening agents include sucrose, lactose, mannitoland artificial sweetening agents such as saccharin, and any number ofspray dried flavours. Flavouring agents include natural flavoursextracted from plants such as fruits and synthetic blends of compoundswhich produce a pleasant sensation, such as, but not limited topeppermint and methyl salicylate. Wetting agents include propyleneglycol monostearate, sorbitan monooleate, diethylene glycol monolaurateand polyoxyethylene laural ether. Emetic-coatings include fatty acids,fats, waxes, shellac, ammoniated shellac and cellulose acetatephthalates. Film coatings include hydroxyethylcellulose, sodiumcarboxymethylcellulose, polyethylene glycol 4000 and cellulose acetatephthalate.

The compound, or pharmaceutically acceptable derivative thereof, couldbe provided in a composition that protects it from the acidicenvironment of the stomach. For example, the composition can beformulated in an enteric coating that maintains its integrity in thestomach and releases the active compound in the intestine. Thecomposition may also be formulated in combination with an antacid orother such ingredient.

When the dosage unit form is a capsule, it can contain, in addition tomaterial of the above type, a liquid carrier such as a fatty oil. Inaddition, dosage unit forms can contain various other materials whichmodify the physical form of the dosage unit, for example, coatings ofsugar and other enteric agents. The compounds can also be administeredas a component of an elixir, suspension, syrup, wafer, sprinkle, chewinggum or the like. A syrup may contain, in addition to the activecompounds, sucrose as a sweetening agent and certain preservatives, dyesand colourings and flavours.

The active materials can also be mixed with other active materials whichdo not impair the desired action, or with materials that supplement thedesired action, such as antacids, H2 blockers, and diuretics. The activeingredient is a compound or pharmaceutically acceptable derivativethereof as described herein. Higher concentrations, up to about 98% byweight of the active ingredient may be included.

In all embodiments, tablets and capsules formulations may be coated asknown by those of skill in the art in order to modify or sustaindissolution of the active ingredient. Thus, for example, they may becoated with a conventional enterically digestible coating, such asphenylsalicylate, waxes and cellulose acetate phthalate.

Liquid Compositions for Oral Administration

Liquid oral dosage forms include aqueous solutions, emulsions,suspensions, solutions and/or suspensions reconstituted fromnon-effervescent granules and effervescent preparations reconstitutedfrom effervescent granules. Aqueous solutions include, for example,elixirs and syrups. Emulsions are either oil-in-water or water-in-oil.

Liquid pharmaceutically administrable compositions can, for example, beprepared by dissolving, dispersing, or otherwise mixing an activecompound as defined above and optional pharmaceutical adjuvants in acarrier, such as, for example, water, saline, aqueous dextrose,glycerol, glycols, ethanol, and the like, to thereby form a solution orsuspension. If desired, the pharmaceutical composition to beadministered may also contain minor amounts of nontoxic auxiliarysubstances such as wetting agents, emulsifying agents, solubilizingagents, pH buffering agents and the like, for example, acetate, sodiumcitrate, cyclodextrine derivatives, sorbitan monolaurate,triethanolamine sodium acetate, triethanolamine oleate, and other suchagents.

Elixirs are clear, sweetened, hydroalcoholic preparations.Pharmaceutically acceptable carriers used in elixirs include solvents.Syrups are concentrated aqueous solutions of a sugar, for example,sucrose, and may contain a preservative. An emulsion is a two-phasesystem in which one liquid is dispersed in the form of small globulesthroughout another liquid. Pharmaceutically acceptable carriers used inemulsions are non-aqueous liquids, emulsifying agents and preservatives.Suspensions use pharmaceutically acceptable suspending agents andpreservatives. Pharmaceutically acceptable substances used innon-effervescent granules, to be reconstituted into a liquid oral dosageform, include diluents, sweeteners and wetting agents. Pharmaceuticallyacceptable substances used in effervescent granules, to be reconstitutedinto a liquid oral dosage form, include organic acids and a source ofcarbon dioxide. Colouring and flavouring agents are used in all of theabove dosage forms.

Solvents include glycerin, sorbitol, ethyl alcohol and syrup. Examplesof preservatives include glycerin, methyl and propylparaben, benzoicacid, sodium benzoate and alcohol. Examples of non-aqueous liquidsutilized in emulsions include mineral oil and cottonseed oil. Examplesof emulsifying agents include gelatin, acacia, tragacanth, bentonite,and surfactants such as polyoxyethylene sorbitan monooleate. Suspendingagents include sodium carboxymethylcellulose, pectin, tragacanth, Veegumand acacia. Sweetening agents include sucrose, syrups, glycerin andartificial sweetening agents such as saccharin. Wetting agents includepropylene glycol monostearate, sorbitan monooleate, diethylene glycolmonolaurate and polyoxyethylene lauryl ether. Organic acids includecitric and tartaric acid. Sources of carbon dioxide include sodiumbicarbonate and sodium carbonate. Colouring agents include any of theapproved certified water soluble FD and C dyes, and mixtures thereof.Flavouring agents include natural flavours extracted from plants suchfruits, and synthetic blends of compounds which produce a pleasant tastesensation.

For a solid dosage form, the solution or suspension, in for examplepropylene carbonate, vegetable oils or triglycerides, is in oneembodiment encapsulated in a gelatin capsule. Such solutions, and thepreparation and encapsulation thereof, are disclosed in U.S. Pat. Nos.4,328,245; 4,409,239; and 4,410,545. For a liquid dosage form, thesolution, e.g., for example, in a polyethylene glycol, may be dilutedwith a sufficient quantity of a pharmaceutically acceptable liquidcarrier, e.g., water, to be easily measured for administration.

Alternatively, liquid or semi-solid oral formulations may be prepared bydissolving or dispersing the active compound or salt in vegetable oils,glycols, triglycerides, propylene glycol esters (e.g., propylenecarbonate) and other such carriers, and encapsulating these solutions orsuspensions in hard or soft gelatin capsule shells. Other usefulformulations include those set forth in U.S. Pat. Nos. RE28,819 and4,358,603. Briefly, such formulations include, but are not limited to,those containing a compound provided herein, a dialkylated mono- orpoly-alkylene glycol, including, but not limited to,1,2-dimethoxymethane, diglyme, triglyme, tetraglyme, polyethyleneglycol-350-dimethyl ether, polyethylene glycol-550-dimethyl ether,polyethylene glycol-750-dimethyl ether wherein 350, 550 and 750 refer tothe approximate average molecular weight of the polyethylene glycol, andone or more antioxidants, such as butylated hydroxytoluene (BHT),butylated hydroxyanisole (BHA), propyl gallate, vitamin E, hydroquinone,hydroxycoumarins, ethanolamine, lecithin, cephalin, ascorbic acid, malicacid, sorbitol, phosphoric acid, thiodipropionic acid and its esters,and dithiocarbamates.

Other formulations include, but are not limited to, aqueous alcoholicsolutions including a pharmaceutically acceptable acetal. Alcohols usedin these formulations are any pharmaceutically acceptable water-misciblesolvents having one or more hydroxyl groups, including, but not limitedto, propylene glycol and ethanol. Acetals include, but are not limitedto, di(lower alkyl) acetals of lower alkyl aldehydes such asacetaldehyde diethyl acetal.

Injectables, Solutions and Emulsions

Parenteral administration, in one embodiment characterized by injection,either subcutaneously, intramuscularly or intravenously is alsocontemplated herein. Injectables can be prepared in conventional forms,either as liquid solutions or suspensions, solid forms suitable forsolution or suspension in liquid prior to injection, or as emulsions.The injectables, solutions and emulsions also contain one or moreexcipients. Suitable excipients are, for example, water, saline,dextrose, glycerol or ethanol. In addition, if desired, thepharmaceutical compositions to be administered may also contain minoramounts of non-toxic auxiliary substances such as wetting or emulsifyingagents, pH buffering agents, stabilizers, solubility enhancers, andother such agents, such as for example, sodium acetate, sorbitanmonolaurate, triethanolamine oleate and cyclodextrins.

Implantation of a slow-release or sustained-release system, such that aconstant level of dosage is maintained (see, e.g., U.S. Pat. No.3,710,795) is also contemplated herein. Briefly, a compound providedherein is dispersed in a solid inner matrix, e.g.,polymethylmethacrylate, polybutylmethacrylate, plasticized orunplasticized polyvinylchloride, plasticized nylon, plasticizedpolyethyleneterephthalate, natural rubber, polyisoprene,polyisobutylene, polybutadiene, polyethylene, ethylene-vinylacetatecopolymers, silicone rubbers, polydimethylsiloxanes, silicone carbonatecopolymers, hydrophilic polymers such as hydrogels of esters of acrylicand methacrylic acid, collagen, cross-linked polyvinylalcohol andcross-linked partially hydrolyzed polyvinyl acetate, that is surroundedby an outer polymeric membrane, e.g., polyethylene, polypropylene,ethylene/propylene copolymers, ethylene/ethyl acrylate copolymers,ethylene/vinylacetate copolymers, silicone rubbers, polydimethylsiloxanes, neoprene rubber, chlorinated polyethylene, polyvinylchloride,vinylchloride copolymers with vinyl acetate, vinylidene chloride,ethylene and propylene, ionomer polyethylene terephthalate, butyl rubberepichlorohydrin rubbers, ethylene/vinyl alcohol copolymer,ethylene/vinyl acetate/vinyl alcohol terpolymer, andethylene/vinyloxyethanol copolymer, that is insoluble in body fluids.The compound diffuses through the outer polymeric membrane in a releaserate controlling step. The percentage of active compound contained insuch parenteral compositions is highly dependent on the specific naturethereof, as well as the activity of the compound and the needs of thesubject.

Parenteral administration of the compositions includes intravenous,subcutaneous and intramuscular administrations. Preparations forparenteral administration include sterile solutions ready for injection,sterile dry soluble products, such as lyophilized powders, ready to becombined with a solvent just prior to use, including hypodermic tablets,sterile suspensions ready for injection, sterile dry insoluble productsready to be combined with a vehicle just prior to use and sterileemulsions. The solutions may be either aqueous or nonaqueous.

If administered intravenously, suitable carriers include physiologicalsaline or phosphate buffered saline (PBS), and solutions containingthickening and solubilizing agents, such as glucose, polyethyleneglycol, and polypropylene glycol and mixtures thereof.

Pharmaceutically acceptable carriers used in parenteral preparationsinclude aqueous vehicles, nonaqueous vehicles, antimicrobial agents,isotonic agents, buffers, antioxidants, local anesthetics, suspendingand dispersing agents, emulsifying agents, sequestering or chelatingagents and other pharmaceutically acceptable substances.

Examples of aqueous vehicles include Sodium Chloride Injection, RingersInjection, Isotonic Dextrose Injection, Sterile Water Injection,Dextrose and Lactated Ringers Injection. Nonaqueous parenteral vehiclesinclude fixed oils of vegetable origin, cottonseed oil, corn oil, sesameoil and peanut oil. Antimicrobial agents in bacteriostatic orfungistatic concentrations must be added to parenteral preparationspackaged in multiple-dose containers which include phenols or cresols,mercurials, benzyl alcohol, chlorobutanol, methyl and propylp-hydroxybenzoic acid esters, thimerosal, benzalkonium chloride andbenzethonium chloride. Isotonic agents include sodium chloride anddextrose. Buffers include phosphate and citrate. Antioxidants includesodium bisulfate. Local anesthetics include procaine hydrochloride.Suspending and dispersing agents include sodium carboxymethylcelluose,hydroxypropyl methylcellulose and polyvinylpyrrolidone. Emulsifyingagents include Polysorbate 80 (TWEEN® 80). A sequestering or chelatingagent of metal ions including EDTA. Pharmaceutical carriers also includeethyl alcohol, polyethylene glycol and propylene glycol for watermiscible vehicles; and sodium hydroxide, hydrochloric acid, citric acidor lactic acid for pH adjustment.

The concentration of the pharmaceutically active compound is adjusted sothat an injection provides an effective amount to produce the desiredpharmacological effect. The exact dose depends on the age, weight andcondition of the patient or animal as is known in the art.

The unit-dose parenteral preparations are packaged in an ampoule, a vialor a syringe with a needle. All preparations for parenteraladministration must be sterile, as is known and practiced in the art.

Illustratively, intravenous or intra-arterial infusion of a sterileaqueous solution containing an active compound is an effective mode ofadministration. Another embodiment is a sterile aqueous or oily solutionor suspension containing an active material injected as necessary toproduce the desired pharmacological effect.

Injectables are designed for local and systemic administration. In oneembodiment, a therapeutically effective dosage is formulated to containa concentration of at least about 0.1% w/w up to about 90% w/w or more,in certain embodiments more than 1% w/w of the active compound to thetreated tissue(s).

The compound may be suspended in micronized or other suitable form ormay be derivatized to produce a more soluble active product or toproduce a prodrug. The form of the resulting mixture depends upon anumber of factors, including the intended mode of administration and thesolubility of the compound in the selected carrier or vehicle. Theeffective concentration is sufficient for ameliorating the symptoms ofthe condition and may be empirically determined.

Lyophilized Powders

Of interest herein are also lyophilized powders, which can bereconstituted for administration as solutions, emulsions and othermixtures. They may also be reconstituted and formulated as solids orgels.

The sterile, lyophilized powder is prepared by dissolving a compoundprovided herein, or a pharmaceutically acceptable derivative thereof, ina suitable solvent. The solvent may contain an excipient which improvesthe stability or other pharmacological component of the powder orreconstituted solution, prepared from the powder. Excipients that may beused include, but are not limited to, dextrose, sorbital, fructose, cornsyrup, xylitol, glycerin, glucose, sucrose or other suitable agent. Thesolvent may also contain a buffer, such as citrate, sodium or potassiumphosphate or other such buffer known to those of skill in the art at, inone embodiment, about neutral pH. Subsequent sterile filtration of thesolution followed by lyophilization under standard conditions known tothose of skill in the art provides the desired formulation. In oneembodiment, the resulting solution will be apportioned into vials forlyophilization. Each vial will contain a single dosage or multipledosages of the compound. The lyophilized powder can be stored underappropriate conditions, such as at about 4° C. to room temperature.

Reconstitution of this lyophilized powder with water for injectionprovides a formulation for use in parenteral administration. Forreconstitution, the lyophilized powder is added to sterile water orother suitable carrier. The precise amount depends upon the selectedcompound. Such amount can be empirically determined.

Topical Administration

Topical mixtures are prepared as described for the local and systemicadministration. The resulting mixture may be a solution, suspension,emulsions or the like and are formulated as creams, gels, ointments,emulsions, solutions, elixirs, lotions, suspensions, tinctures, pastes,foams, aerosols, irrigations, sprays, suppositories, bandages, dermalpatches or any other formulations suitable for topical administration.

The compounds or pharmaceutically acceptable derivatives thereof may beformulated as aerosols for topical application, such as by inhalation(see, e.g., U.S. Pat. Nos. 4,044,126, 4,414,209, and 4,364,923, whichdescribe aerosols for delivery of a steroid useful for treatment ofinflammatory diseases, particularly asthma). These formulations foradministration to the respiratory tract can be in the form of an aerosolor solution for a nebulizer, or as a microfine powder for insufflation,alone or in combination with an inert carrier such as lactose. In such acase, the particles of the formulation will, in one embodiment, havediameters of less than 50 microns, in one embodiment less than 10microns.

The compounds may be formulated for local or topical application, suchas for topical application to the skin and mucous membranes, such as inthe eye, in the form of gels, creams, and lotions and for application tothe eye or for intracisternal or intraspinal application. Topicaladministration is contemplated for transdermal delivery and also foradministration to the eyes or mucosa, or for inhalation therapies. Nasalsolutions of the active compound alone or in combination with otherpharmaceutically acceptable excipients can also be administered.

These solutions, particularly those intended for ophthalmic use, may beformulated as 0.01%-10% (vol %) isotonic solutions, pH about 5-7, withappropriate salts.

Compositions for Other Routes of Administration

Other routes of administration, such as transdermal patches, includingiontophoretic and electrophoretic devices, and rectal administration,are also contemplated herein.

Transdermal patches, including iontophoretic and electrophoreticdevices, are well known to those of skill in the art. For example, suchpatches are disclosed in U.S. Pat. Nos. 6,267,983, 6,261,595, 6,256,533,6,167,301, 6,024,975, 6,010715, 5,985,317, 5,983,134, 5,948,433, and5,860,957.

For example, pharmaceutical dosage forms for rectal administration arerectal suppositories, capsules and tablets for systemic effect. Rectalsuppositories are used herein mean solid bodies for insertion into therectum which melt or soften at body temperature releasing one or morepharmacologically or therapeutically active ingredients.Pharmaceutically acceptable substances utilized in rectal suppositoriesare bases or vehicles and agents to raise the melting point. Examples ofbases include cocoa butter (theobroma oil), glycerin-gelatin, carbowax(polyoxyethylene glycol) and appropriate mixtures of mono-, di- andtriglycerides of fatty acids. Combinations of the various bases may beused. Agents to raise the melting point of suppositories includespermaceti and wax. Rectal suppositories may be prepared either by thecompressed method or by molding. The weight of a rectal suppository, inone embodiment, is about 2 to 3 gm.

Tablets and capsules for rectal administration are manufactured usingthe same pharmaceutically acceptable substance and by the same methodsas for formulations for oral administration.

Targeted Formulations

The compounds provided herein, or pharmaceutically acceptablederivatives thereof, may also be formulated to be targeted to aparticular tissue, receptor, or other area of the body of the subject tobe treated. Many such targeting methods are well known to those of skillin the art. All such targeting methods are contemplated herein for usein the instant compositions. For non-limiting examples of targetingmethods, see, e.g., U.S. Pat. Nos. 6,316,652, 6,274,552, 6,271,359,6,253,872, 6,139,865, 6,131,570, 6,120,751, 6,071,495, 6,060,082,6,048,736, 6,039,975, 6,004,534, 5,985,307, 5,972,366, 5,900,252,5,840,674, 5,759,542 and 5,709,874.

In one embodiment, liposomal suspensions, including tissue-targetedliposomes, such as tumor-targeted liposomes, may also be suitable aspharmaceutically acceptable carriers. These may be prepared according tomethods known to those skilled in the art. For example, liposomeformulations may be prepared as described in U.S. Pat. No. 4,522,811.Briefly, liposomes such as multilamellar vesicles (MLV's) may be formedby drying down egg phosphatidyl choline and brain phosphatidyl serine(7:3 molar ratio) on the inside of a flask. A solution of a compoundprovided herein in phosphate buffered saline lacking divalent cations(PBS) is added and the flask shaken until the lipid film is dispersed.The resulting vesicles are washed to remove unencapsulated compound,pelleted by centrifugation, and then resuspended in PBS.

Co-Administration with Other Drugs

In accordance with another aspect of the present invention, it iscontemplated that compounds as described herein may be administered to asubject in need thereof in combination with medication considered bythose of skill in the art to be current standard of care for thecondition of interest. Such combinations provide one or more advantagesto the subject, e.g., requiring reduced dosages to achieve similarbenefit, obtaining the desired palliative affect in less time, and thelike.

Compounds in accordance with the present invention may be administeredas part of a therapeutic regimen with other drugs. It may desirable toadminister a combination of active compounds, for example, for thepurpose of treating a particular disease or condition. Accordingly, itis within the scope of the present invention that two or morepharmaceutical compositions, at least one of which contains a compoundof Formula (I) according to the present invention, may be combined inthe form of a kit suitable for co-administration of the compositions.

In one embodiment of the methods of the present inventions a compound ofFormula I may be administered with a second therapeutic agent. In oneembodiment the second therapeutic agent is selected from the groupconsisting of an anti-cancer agent, an anti-inflammatory agent, ananti-hypertensive agent, an anti-fibrotic agent, an anti-angiogenicagent, an anti-diabetic agent, and an immunosuppressive agent.

When two or more active ingredients are co-administered, the activeingredients may be administered simultaneously, sequentially orseparately. In one embodiment the compound of Formula I isco-administered simultaneously with a second therapeutic agent. Inanother embodiment the compound of Formula I and the second therapeuticagent are administered sequentially. In a further embodiment thecompound of Formula I and the second therapeutic agent are administeredseparately.

The invention will now be described in greater detail with reference tothe following non-limiting examples. The examples are intended to serveto illustrate the invention and should not be construed as limiting thegenerality of the disclosure of the description throughout thisspecification.

Example 1 Preparation of the synthons (Z)-tert-butyl2-(bromomethyl)-3-fluoroallylcarbamate and (E)-tert-butyl2-(bromomethyl)-3-fluoroallylcarbamate

Preparation of tert-butyl3-(tert-butyldimethylsilyloxy)-2-hydroxypropylcarbamate

To a stirred solution of 3-amino-1,2-propanediol (10.0 g, 0.11 mol) andtriethylamine (23 mL, 0.17 mol) in MeOH (200 mL) at room temperature wasadded di-tert-butyl dicarbonate (26.4 g, 0.12 mol). The resultingsolution was left to stir at room temperature overnight. The reactionmixture was concentrated under reduced pressure then co-evaporated withtoluene to remove all the MeOH. The crude residue was taken up in CH₂Cl₂and, after cooling to 0° C., imidazole andtert-butyl-(chloro)dimethylsilane were sequentially added. The resultingmixture was left to stir at this temperature for 2 h. The reactionmixture was partitioned between water (100 mL) and CH₂Cl₂ (70 mL) andthe aqueous layer was extracted with further CH₂Cl₂ (2×70 mL). Thecombined organics were dried over Na₂SO₄ and concentrated in vacuo. Thecrude residue was purified over silica gel eluting with n-hexanefollowed by 10% ethyl acetate in hexanes to afford tert-butyl3-(tert-butyldimethylsilyloxy)-2-hydroxypropylcarbamate (32.6 g, 97.3%)as a colourless oil. ¹H-NMR (300 MHz, CDCl₃) δppm: 0.09 (6H, s), 0.91(9H, s), 1.46 (9H, s), 2.86 (1H, br d, J 4.2 Hz), 3.13 (1H, ddd, J 14.1,6.7, 5.3 Hz), 3.30-3.43 (1H, m), 3.54 (1H, dd, J 10.1, 6.2 Hz), 3.66(1H, dd, J 10.1, 4.5 Hz), 3.70-3.80 (1H, m), 4.98 (1H, br s).

Preparation of tert-butyl3-(tert-butyldimethylsilyloxy)-2-oxopropylcarbamate

To a stirring solution of oxalyl chloride (13.6 mL, 0.16 mol) in dryCH₂Cl₂ (150 mL) at −78° C. under N₂ was added DMSO (15.2 mL, 0.21 mol)dropwise over 30 min. After complete addition the resulting solution wasstirred at −78° C. for 1 h. A solution of tert-butyl3-(tert-butyldimethyl-silyloxy)-2-hydroxypropylcarbamate (32.6 g, 0.11mol) in CH₂Cl₂ (50 mL) was then added dropwise over 20 min. Stirring wascontinued for a further 1 hour at which time triethylamine (59.6 mL,0.43 mol) was added. The cooling bath was removed and the reactionmixture was allowed to warm to room temperature. The reaction mixturewas partitioned between water (100 mL) and CH₂Cl₂ (70 mL) and theaqueous layer was extracted with further CH₂Cl₂ (2×70 mL); the combinedorganics were dried over Na₂SO₄ and concentrated under a stream ofnitrogen gas. The crude residue was purified over silica gel elutingwith 5% ethylacetate in n-hexane to give tert-butyl3-(tert-butyldimethylsilyloxy)-2-oxopropylcarbamate (29.8 g, 92%) as apale yellow oil. ¹H-NMR (300 MHz; CDCl₃) δppm: 0.11 (6H, s), 0.94 (9H,s), 1.47 (9H, s), 3.92 (2H, s), 4.26 (2H, d, J 4.6 Hz), 5.22 (1H, br s).

Preparation of tert-butyl2-((tert-butyldimethylsilyloxy)methyl)-3-fluoroallylcarbamate

To a vigorously stirring suspension offluoromethyl(triphenyl)-phosphonium tetrafluoroborate (18.9 g, 49.4mmol) in dry THF (190 mL) at −20° C. under N₂ was added sodiumbis(trimethylsilyl)amide (1.0 M in THF; 49.4 mL, 49.4 mmol) slowly over10 min. The resulting deep orange solution was left to stir at thistemperature for 15 min. A solution of tert-butyl3-(tert-butyldimethylsilyloxy)-2-oxopropylcarbamate (10.0 g, 33.0 mmol)in THF (10 mL) was then added slowly over 10 min. After completeaddition, stirring was continued for a further 1 h during which time thereaction was allowed to warm slowly to room temperature. The reactionwas quenched by addition of water (5 mL) and the reaction mixture wasconcentrated in vacuo. The residue was partitioned between water (100mL) and diethyl ether (100 mL) and the aqueous layer was extracted withfurther diethyl ether (2×100 ml). The combined organics were dried overNa₂SO₄ and concentrated under reduced pressure. The crude residue waspurified over silica gel eluting with n-hexane followed by 6%ethylacetate in n-hexane to give tert-butyl2-((tert-butyldimethylsilyloxy)methyl)-3-fluoroallylcarbamate as amixture of E/Z double-bond isomers (E/Z=1:1; 9.9 g, 94%). The isomerswere not separated at this stage.

Preparation of (E)-tert-butyl 3-fluoro-2-(hydroxymethyl)allylcarbamateand (Z)-tert-butyl 3-fluoro-2-(hydroxymethyl)allylcarbamate

To a stirring solution of tert-butyl2-((tert-butyldimethylsilyloxy)methyl)-3-fluoroallylcarbamate (E/Z=1:1;12.0 g, 37.6 mmol) in THF (30 mL) at room temperature was added TBAF(1.0 M in THF; 45.1 mL, 45.1 mmol). The resulting solution was left tostir for 30 min. The reaction mixture was partitioned between water (70mL) and ethyl acetate (50 mL). The aqueous layer was extracted withethyl acetate (50 mL) and the combined organics were washed withsaturated aqueous NH₄Cl (70 mL) followed by brine (70 mL). After dryingover Na₂SO₄, the organics were concentrated in vacuo. Purification ofthe crude material over silica gel eluting with 20% ethyl acetate and 5%THF in n-hexane gave (Z)-tert-butyl3-fluoro-2-(hydroxymethyl)-allylcarbamate (0.5 g, 6.5%), (E)-tert-butyl3-fluoro-2-(hydroxymethyl)allylcarbamate (1.2 g, 15.6%) and a mixture ofthe E/Z isomers (5.5 g, 71.4%).

(Z)-tert-Butyl 3-fluoro-2-(hydroxymethyl)allylcarbamate: ¹H-NMR (300MHz; CDCl₃) δppm: 1.46 (9H, s), 3.41 (1H, br s), 3.74 (2H, dd, J 6.5,3.1 Hz), 4.28 (2H, dd, J 6.0, 2.3 Hz), 4.87 (1H, br s), 6.53 (1H, dd, J83.5 Hz).

(E)-tert-Butyl 3-fluoro-2-(hydroxymethyl)allylcarbamate: ¹H-NMR (300MHz; CDCl₃) δppm: 1.47 (9H, s), 3.78 (1H, t, J 6.4 Hz), 3.93-4.02 (4H,m), 4.94 (1H, br s), 6.63 (1H, d, J 83.6 Hz).

Preparation of (Z)-tert-butyl 2-(bromomethyl)-3-fluoroallylcarbamate

To a stirring solution of (Z)-tert-butyl3-fluoro-2-(hydroxymethyl)-allylcarbamate (0.50 g, 2.44 mmol) in acetone(15 mL) at 0° C. under N₂ was added sequentially triethylamine (0.51 mL,3.65 mmol) and methanesulfonyl chloride (0.23 mL, 2.92 mmol). Theresulting mixture was stirred at this temperature for 30 min. Thereaction mixture was filtered to remove the precipitated salts and thefilter cake was washed with further acetone (10 mL). The filtrate wascharged with lithium bromide (1.06 g, 12.18 mmol) and the resultingsuspension was stirred at room temperature for 1 h. The reaction mixturewas partitioned between water (25 mL) and ethyl acetate (25 mL) and theaqueous layer was extracted with further ethyl acetate (25 mL). Thecombined organics were washed with brine (25 mL), dried over Na₂SO₄ andconcentrated in vacuo to give (Z)-tert-butyl2-(bromomethyl)-3-fluoroallylcarbamate as a pale yellow oil (0.63 g,96%). ¹H-NMR (300 MHz; CDCl₃) δppm: 1.47 (9H, s), 3.80 (2H, br s), 4.09(2H, d, J 2.6 Hz), 4.75 (1H, br s), 6.65 (1H, d, J 81.9 Hz).

Preparation of (E)-tert-butyl 2-(bromomethyl)-3-fluoroallylcarbamate

To a stirring solution of (E)-tert-butyl3-fluoro-2-(hydroxymethyl)-allylcarbamate (1.20 g, 5.85 mmol) in acetone(20 mL) at 0° C. under N₂ was added sequentially triethylamine (1.22 mL,8.77 mmol) and methanesulfonyl chloride (0.54 mL, 7.02 mmol). Theresulting mixture was stirred at this temperature for 30 min. Thereaction mixture was filtered to remove the precipitated salts and thefilter cake was washed with further acetone (10 mL). The filtrate wascharged with lithium bromide (2.54 g, 29.24 mmol) and the resultingsuspension was stirred at room temperature for 1 h. The reaction mixturewas partitioned between water (25 mL) and ethyl acetate (25 mL) and theaqueous layer was extracted with further ethyl acetate (25 mL). Thecombined organics were washed with brine (25 mL), dried over Na₂SO₄ andconcentrated in vacuo to give (E)-tert-butyl2-(bromomethyl)-3-fluoroallylcarbamate as a pale yellow oil (1.46 g,93%). ¹H-NMR (300 MHz; CDCl₃) δppm: 1.47 (9H, s), 3.97 (2H, dd, J 3.5,0.7 Hz), 4.02 (2H, br d, J 6.1 Hz), 4.78 (1H, br s), 6.79 (1H, d, J 81.1Hz).

Example 2 Procedure A: Preparation of (Z)-tert-butyl2-((4-(dimethylcarbamoyl)phenoxy)-methyl)-3-fluoroallylcarbamate

To a vigorously stirring suspension of (Z)-tert-butyl2-(bromomethyl)-3-fluoroallylcarbamate (430.0 mg, 1.60 mmol) andpotassium carbonate (332.5 mg, 2.41 mmol) in dry DMF (2.0 mL) at roomtemperature under N₂ was added 4-hydroxy-N,N-dimethylbenzamide (291.4mg, 1.76 mmol). The resulting mixture was stirred at room temperatureovernight. The reaction mixture was partitioned between water (40 mL)and ethyl acetate (20 mL) and the aqueous layer was extracted withfurther ethyl acetate (2×20 ml). The combined organics were washed withsaturated aqueous NH₄Cl (40 mL), brine (40 mL), dried over Na₂SO₄ andconcentrated under reduced pressure. Purification of the crude materialover silica gel eluting with 60% ethyl acetate in n-hexane followed by75% ethyl acetate in n-hexane gave (Z)-tert-butyl2-((4-(dimethylcarbamoyl)phenoxy)methyl)-3-fluoroallylcarbamate (520.0mg, 92%) as a colourless oil. ¹H-NMR (300 MHz; CDCl₃) δppm: 1.44 (9H,s), 3.07 (6H, br s), 3.78 (2H, br s), 4.74 (2H, dd, J 2.7, 0.8 Hz), 4.80(1H, br s), 6.75 (1H, d, J 82.7 Hz), 6.95 (2H, d, J 8.9 Hz), 7.42 (2H,d, J 8.8 Hz).

Procedure B: Preparation of(Z)-4-(2-(aminomethyl)-3-fluoroallyloxy)-N,N-dimethyl-benzamidehydrochloride (Compound 18)

To a stirring solution of (Z)-tert-butyl2-((4-(dimethylcarbamoyl)-phenoxy)methyl)-3-fluoroallylcarbamate (520.0mg, 1.48 mmol) in CH₂Cl₂ (8.0 mL) at room temperature was addedtrifluoroacetic acid (2.0 mL). The resulting mixture was stirred at roomtemperature for 30 min. All volatiles were removed in vacuo and theresidue was co-evaporated with CH₂Cl₂ (2×20 mL) to removetrifluoroacetic acid. The resulting oil was taken up in ethyl acetate(3.0 mL) and then ethereal HCl (2.0 M in diethyl ether; 1.0 mL, 2.0mmol) was added. The precipitate formed was isolated and dried underreduced pressure to afford(Z)-4-(2-(aminomethyl)-3-fluoroallyloxy)-N,N-dimethylbenzamidehydrochloride (301 mg, 71%) as a pale yellow solid; m.p.=135-137° C.;¹H-NMR (300 MHz; MeOD) δppm: 3.06 (3H, br s), 3.10 (3H, br s), 3.71 (2H,d, J 3.0 Hz), 4.88 (2H, dd, J 2.8, 0.8 Hz), 7.11 (2H, d, J 8.9 Hz), 7.13(1H, d, J 80.8 Hz), 7.45 (2H, d, J 8.9 Hz).

Procedure C: Preparation of (Z)-tert-butyl2-((4-(N,N-dimethylsulfamoyl)phenoxy)methyl)-3-fluoroallylcarbamate

To a vigorously stirring suspension of (Z)-tert-butyl2-(bromomethyl)-3-fluoroallylcarbamate (232.0 mg, 0.87 mmol) in dry DMF(2.0 mL) at room temperature under N₂ was sequentially added potassiumcarbonate (300.0 mg, 2.16 mmol) and 4-hydroxy-N,N-dimethylbenzamide(174.0 mg, 0.87 mmol). The resulting suspension was left to stir at roomtemperature for 2 h. The reaction mixture was partitioned betweensaturated aqueous NH₄Cl (40 mL) and ethyl acetate (20 mL) and theaqueous layer was extracted with further ethyl acetate (20 ml). Thecombined organics dried over Na₂SO₄ and concentrated under reducedpressure. Purification of the crude material over silica gel elutingwith 50% ethyl acetate in n-hexane gave (Z)-tert-butyl2-((4-(N,N-dimethylsulfamoyl)phenoxy)methyl)-3-fluoroallylcarbamate(279.0 mg, 83%) as a colourless oil. ¹H-NMR (300 MHz; CDCl₃) δppm: 1.42(9H, s), 2.69 (6H, s), 3.79 (2H, br s), 4.76 (2H, d, J 2.7 Hz), 4.81(1H, br s), 6.76 (1H, d, J 82.6 Hz), 7.04 (2H, d, J 8.9 Hz), 7.72 (2H,d, J 9.0 Hz).

Procedure D: Preparation of(Z)-4-(2-(aminomethyl)-3-fluoroallyloxy)-N,N-dimethyl-benzenesulfonamideHydrochloride (Compound 10)

To a stirring solution of (Z)-tert-butyl2-((4-(N,N-dimethylsulfamoyl)phenoxy)methyl)-3-fluoroallylcarbamate(279.0 mg, 0.72 mmol) in CH₂Cl₂ (4.0 mL) at room temperature was addedtrifluoroacetic acid (1.0 mL). The resulting mixture was stirred at roomtemperature for 30 min. All volatiles were removed in vacuo and theresidue was co-evaporated with CH₂Cl₂ (2×20 mL). The resulting oil wastaken up in ethyl acetate/MeOH (5:1; 3.0 mL) and then ethereal HCl (2.0M in diethyl ether; 0.5 mL, 1.0 mmol) was added. The precipitate formedwas isolated and dried under reduced pressure to afford(Z)-4-(2-(aminomethyl)-3-fluoroallyloxy)-N,N-dimethylbenzenesulfonamidehydrochloride (196.0 mg, 84%) as a white solid; m.p. 185-187° C.; ¹H-NMR(300 MHz; d₆-DMSO) δppm: 3.39 (6H, br s), 3.54 (2H, br s), 4.81 (2H, d,J 2.3 Hz), 7.16 (2H, d, J 9.0 Hz), 7.24 (1H, d, J 82.3 Hz), 7.25 (2H, brs), 7.77 (2H, d, J 9.0 Hz).

Example 3

The following compounds were prepared according to procedures A and B asset forth in Example 2.

(Z)-4-(2-(Aminomethyl)-3-fluoroallyloxy)-N-tert-butylbenzamideHydrochloride (Compound 1)

Beige solid; m.p. 180-184° C.; ¹H-NMR (300 MHz; CD₃OD) δ ppm: 1.45 (9H,s), 3.70 (2H, d, J 2.2 Hz), 4.86 (2H, dd, J 2.9, 0.7 Hz), 7.06 (2H, d, J9.0 Hz), 7.13 (1H, d, J 80.9 Hz), 7.76 (2H, d, J 8.9 Hz).

(Z)-4-(2-(Aminomethyl)-3-fluoroallyloxy)-3-fluoro-N,N-dimethyl-benzamideHydrochloride (Compound 4)

Brown solid; ¹H-NMR (300 MHz; CD₃OD) δ ppm: 3.04 (3H, br s), 3.09 (3H,br s), 3.73 (2H, d, J 2.4 Hz), 4.93 (2H, dd, J 2.9, 0.8 Hz), 7.16 (1H, dJ 90.0 Hz), 7.25-7.29 (2H, m)

(Z)-4-(2-(Aminomethyl)-3-fluoroallyloxy)-3-chloro-N,N-dimethyl-benzamideHydrochloride (Compound 6)

Brown solid; ¹H-NMR (300 MHz; CD₃OD) δ ppm: 3.04 (3H, br s), 3.09 (3H,br s), 3.76 (2H, d, J 2.3 Hz), 4.96 (2H, dd, J 2.8, 0.9 Hz), 7.16 (1H,d, 80.6 Hz), 7.26 (1H, d, J 8.6 Hz), 7.43 (1H, dd, J 8.5, 2.1 Hz), 7.55(1H, d, J 2.0 Hz)

(Z)-4-(2-(Aminomethyl)-3-fluoroallyloxy)-3-bromo-N,N-dimethyl-benzamideHydrochloride (Compound 20)

Beige-coloured solid; m.p. 54-57° C.; ¹H-NMR (300 MHz; CD₃OD) δ ppm:3.04 (3H, br s), 3.09 (3H, br s), 3.78 (2H, d, J 2.4 Hz), 4.95 (2H, dd,J 2.9, 0.9 Hz), 7.15 (1H, d, J 80.5 Hz), 7.22 (1H, d, J 8.5 Hz), 7.47(1H, dd, J 8.5, 2.1 Hz), 7.71 (1H, d, J 2.0 Hz)

4-(2-(Aminomethyl)-3-fluoroallylthio)-N,N-dimethylbenzamideHydrochloride as a Mixture of E and Z Isomers (Compounds 8E and 8Z)

Colorless solid; ¹H-NMR (300 MHz; CD₃OD) δ ppm: 2.99 (3H, br s), 3.00(3H, br s), 3.10 (6H, br s), 3.64 (2H, d, J 3.0 Hz), 3.71 (2H, dd, J3.1, 1.1 Hz), 3.77 (2H, d, J 1.0 Hz), 3.87 (2H, dd, J 2.1, 0.8 Hz), 6.82(1H, d, J 82.1 Hz), 6.93 (1H, d, J 81.6 Hz), 7.38 (2H, d, J 8.6 Hz),7.41 (2H, d, J 8.6 Hz), 7.48 (2H, d, J 8.6 Hz), 7.49 (2H, d, J 8.3 Hz).

(E)-4-(2-(Aminomethyl)-3-fluoroallyloxy)-N-isopropylbenzamideTrifluoroacetate (Compound 39)

Yellow gum; ¹H-NMR (300 MHz; d₆-DMSO) δppm: 1.13 (6H, d, J 6.9 Hz), 3.58(2H, d, J 5.1 Hz), 4.05 (1H, septet, J 6.6 Hz), 4.65 (2H, d, J 3.6 Hz),7.02 (2H, d, J 6.9 Hz), 7.32 (1H, d, J 81.9 Hz), 7.82 (2H, d, J 6.9 Hz),8.07 (1H, d, J 7.5 Hz), 8.18 (3H, br s).

(E)-4-(2-(Aminomethyl)-3-fluoroallyloxy)-N-tert-butylbenzamideHydrochloride (Compound 23)

Colorless powder; m.p. 140-142° C.; ¹H-NMR (300 MHz; d₆-DMSO) δppm: 1.37(9H, s), 3.60 (2H, d, J 3.9 Hz), 4.68 (2H, d, J 3.6 Hz), 7.02 (2H, d, J6.9 Hz), 7.34 (1H, d, J 82.5 Hz), 7.61 (1H, s), 7.81 (2H, d, J 6.9 Hz),8.28 (3H, br s).

(E)-4-(2-(Aminomethyl)-3-fluoroallyloxy)-N,N-diethylbenzamideHydrochloride (Compound 24)

Brown solid; ¹H-NMR (300 MHz; CD₃OD) δ ppm: 1.18 (3H, br s), 1.25 (3H,br s), 3.37 (2H, br s), 3.56 (2H, br s), 3.83 (2H, s), 4.68 (2H, d, J3.5 Hz), 7.12 (2H, d, J 8.6 Hz), 7.40 (2H, d, J 8.7 Hz).

(E)-4-(2-(Aminomethyl)-3-fluoroallyloxy)-N-methylbenzamide Hydrochloride(Compound 25)

Colorless solid; m.p. 203-205° C.; ¹H-NMR (300 MHz; CD₃OD) δ ppm: 2.90(3H, s), 3.83 (2H, d, J 1.8 Hz), 4.67 (2H, dd, J 3.7, 0.8 Hz), 7.07 (2H,d, J 9.0 Hz), 7.24 (1H, d, J 81.2 Hz), 7.81 (2H, d, J 9.0 Hz).

(Z)-4-(2-(Aminomethyl)-3-fluoroallyloxy)benzamide Hydrochloride(Compound 2)

Colorless solid; m.p. 195-198° C.; 1H-NMR (300 MHz; MeOD) δppm: 3.72(2H, d, J 2.2 Hz), 4.90 (2H, dd, J 2.9, 0.8 Hz), 7.11 (2H, d, J 9.0 Hz),7.14 (1H, d, J 80.8 Hz), 7.90 (2H, d, J 9.0 Hz).

(E)-4-(2-(Aminomethyl)-3-fluoroallyloxy)benzamide Hydrochloride(Compound 3)

Colorless solid; m.p. 225-228° C.; 1H-NMR (300 MHz; MeOD) δppm: 3.85(2H, s), 4.70 (2H, dd, J 3.6, 1.0 Hz), 7.10 (2H, d, J 9.0 Hz), 7.26 (1H,d, J 81.2 Hz), 7.90 (2H, d, J 9.0 Hz).

(E)-4-(2-(Aminomethyl)-3-fluoroallyloxy)-N,N-dimethylbenzamideHydrochloride (Compound 13)

m.p. 185-187° C.; ¹H-NMR (300 MHz; d₆-DMSO) δppm: 2.95 (6H, s), 3.60(2H, d (br), J 4.2 Hz), 4.67 (2H, d, J 3.6 Hz), 7.03 (2H, d, J 8.7 Hz),7.33 (1H, d, J 82.2), 7.40 (2H, d, J 8.7 Hz), 8.29 (3H, br s).

(Z)-4-(2-(Aminomethyl)-3-fluoroallyloxy)-N,N,2-trimethylbenzamideHydrochloride (Compound 26)

¹H-NMR (300 MHz; DMSO) δppm: 2.17 (3H, s), 2.75 (3H, s), 2.98 (3H, s),3.54 (2H, m (br)), 4.72 (2H, d, J 2.4 Hz), 6.85 (1H, dd, J 2.4, 8.4 Hz),6.89 (1H, d, J 2.4 Hz), 7.10 (1H, d, J 8.4 Hz), 7.21 (1H, d, J 82.2 Hz),8.15 (3H, s).

4-(2-(Aminomethyl)-3-fluoroallyloxy)-3-methoxy-N,N-dimethylbenzamideHydrochloride as a Mixture of E and Z Isomers (Compounds 7E and 7Z)

E-Isomer

¹H-NMR (300 MHz; DMSO) δppm: 2.95 (6H, s), 3.52 (2H, m (br)), 3.79 (3H,s), 4.65 (2H, d, J 3.3 Hz), 6.95-7.09 (3H, m), 7.24 (1H, d, J 82.0 Hz),8.25 (3H, s).

Z-Isomer

¹H-NMR (300 MHz; DMSO) δppm: 2.95 (6H, s), 3.59 (2H, m (br)), 3.79 (3H,s), 4.77 (2H, d, J 2.1 Hz), 6.95-7.09 (3H, m), 7.29 (1H, d, J 82.0 Hz),8.25 (3H, s).

Example 4

The following compounds were prepared according to procedures C and D asset forth in Example 2.

(E)-4-(2-(Aminomethyl)-3-fluoroallyloxy)benzenesulfonamide Hydrochloride(Compound 11)

Colorless solid; m.p. 107-110° C.; 1H-NMR (300 MHz; MeOD) δppm: 3.85(2H, d, J 2.0 Hz) 4.71 (2H, dd, J 3.6, 0.8 Hz), 7.16 (2H, d, J 9.0 Hz),7.27 (1H, d, J 81.5 Hz), 7.88 (2H, d, J 9.0 Hz).

(E)-4-(2-(Aminomethyl)-3-fluoroallyloxy)-N,N-dimethylbenzenesulfonamideHydrochloride (Compound 14)

m.p. 178-180° C.; ¹H-NMR (300 MHz; d₆-DMSO) δppm: 2.57 (6H, s), 3.61(2H, d (br), J 2.1 Hz), 4.73 (2H, d, J 3.3 Hz), 7.22 (2H, d, J 8.7 Hz),7.36 (1H, d, J 82.2 Hz), 7.71 (2H, d, J 8.7 Hz), 8.29 (3H, brs).

(Z)-3-(2-(Aminomethyl)-3-fluoroallyloxy)-N,N-dimethylbenzenesulfonamideHydrochloride(Compound 15)

Off white solid; m.p. 140-142° C.; ¹H-NMR (300 MHz; CD₃OD) δ ppm: 2.70(6H, s), 3.71 (2H, d, J 2.3 Hz), 4.90 (2H, dd, J 2.9, 0.8 Hz), 7.14 (1H,d, J 80.8 Hz), 7.31-7.62 (4H, m).

(E)-4-(2-(Aminomethyl)-3-fluoroallyloxy)-N-methylbenzenesulfonamideHydrochloride (Compound 28)

Beige solid; m.p. 143-146° C.; 1H-NMR (300 MHz; MeOD) δppm: 2.51 (3H,s), 3.85 (2H, s), 4.73 (2H, d, J 3.3 Hz), 7.19 (2H, d, J 8.8 Hz), 7.27(1H, d, J 81.0 Hz), 7.80 (2H, d, J 8.7 Hz).

(Z)-4-(2-(Aminomethyl)-3-fluoroallyloxy)-N-methylbenzenesulfonamideHydrochloride (Compound 29)

Colorless solid; m.p. 178-180° C.; 1H-NMR (300 MHz; d6-DMSO) δppm: 2.38(3H, d, J 5.0 Hz), 3.55 (2H, br s), 4.81 (2H, d, J 2.3 Hz), 7.20 (2H, d,J 8.9 Hz), 7.25 (1H, d, J 82.0 Hz), 7.34 (1H, q, J 5.1 Hz), 7.73 (2H, d,J 8.9 Hz), 8.15 (3H, br s).

(E)-4-(2-(Aminomethyl)-3-fluoroallyloxy)-N-ethylbenzenesulfonamideHydrochloride (Compound 30)

Colorless solid; m.p. 80-85° C.; 1H-NMR (300 MHz; MeOD) δppm: 1.06 (3H,t, J 7.3 Hz), 2.88 (2H, q, J 7.2 Hz), 3.85 (2H, d, J 2.0 Hz), 4.72 (2H,dd, J 3.6, 0.8 Hz), 7.18 (2H, d, J 9.0 Hz), 7.27 (1H, d, J 81.0 Hz),7.82 (2H, d, J 9.0 Hz).

(Z)-4-(2-(Aminomethyl)-3-fluoroallyloxy)-N-ethylbenzenesulfonamideHydrochloride (Compound 31)

White solid; m.p. 65-67° C.; ¹H-NMR (300 MHz; d₆-DMSO) δppm: 0.96 (3H,t, J 7.2 Hz), 2.74 (2H, dq, J 7.0, 7.2 Hz), 3.55 (2H, br s), 4.80 (2H,br s), 7.19 (2H, d, J 8.8 Hz), 7.25 (1H, d, J 81.9 Hz), 7.44 (1H, t, J5.5 Hz), 7.74 (2H, d, J 8.7 Hz), 8.16 (3H, br s).

(E)-4-(2-(Aminomethyl)-3-fluoroallyloxy)-N-isopropylbenzenesulfonamideHydrochloride (Compound 32)

Colorless solid; m.p. 151-153° C.; 1H-NMR (300 MHz; MeOD) δppm: 1.03(6H, d, J 6.6 Hz), 3.33 (1H, m) 3.85 (2H, s), 4.72 (2H, d, J 3.8 Hz),7.17 (2H, d, J 9.0 Hz), 7.27 (1H, d, J 80.9 Hz), 7.83 (2H, d, J 8.9 Hz).

(Z)-4-(2-(Aminomethyl)-3-fluoroallyloxy)-N-isopropyl-benzenesulfonamideHydrochloride (Compound 33)

White solid; m.p. 50-52° C.; ¹H-NMR (300 MHz; d₆-DMSO) δppm: 0.94 (6H,d, J 6.5 Hz), 3.18 (1H, m), 3.56 (2H, br s), 4.81 (2H, br s), 7.18 (2H,d, J 8.9 Hz), 7.25 (1H, d, J 81.9 Hz), 7.46 (1H, d, J 7.1 Hz), 7.76 (2H,d, J 8.9 Hz), 8.09 (3H, br s).

(Z)-4-(2-(Aminomethyl)-3-fluoroallyloxy)benzenesulfonamide Hydrochloride(Compound 9)

m.p. 227-230° C.; ¹H-NMR (300 MHz; d₆-DMSO) δppm: 3.54 (2H, br), 4.80(2H, s), 7.24 (1H, d, J 82.2 Hz), 7.15 (2H, d, J 8.7 Hz), 7.26 (2H, s),7.77 (2H, d, J 8.7 Hz), 8.14 (3H, br s).

Example 5 Method to Determine the Ability of Compounds of Formula I toInhibit Human Recombinant SSAO/VAP-1

The inhibitory effects of all the compounds of Formula I were testedagainst human recombinant SSAO/VAP-1 using the coupled colorimetricmethod as described for monoamine oxidase, copper-containing amineoxidases and related enzymes (Holt A. and Palcic M., Aperoxidise-coupled continuous absorbance plate-reader assay for flavinmonoamine oxidases, copper-containing amine oxidases and relatedenzymes. Nat. Protoc. 2006, 1, 2498-2505). Briefly, a cloned cDNAtemplate corresponding to residues 34-763 of human SSAO/VAP-1, andincorporating a mouse Ig kappa (κ) signal sequence, N-terminal Flagepitope tag and tobacco etch virus (TEV) cleavage site, was assembled ina mammalian expression vector (pLO-CMV) by Geneart AG. This vectorcontaining human SSAO/VAP-1 residues was transfected into CHO-K1glycosylation mutant cell line, Lec 8. A clone stably expressing humanSSAO/VAP-1 was isolated and cultured in large scale. Active humanSSAO/VAP-1 was purified and recovered using immunoaffinitychromatography. This was used as source for SSAO/VAP-1 activity. Ahigh-throughput colorimetric assay was developed using either 96 or 384well format. Briefly, in a standard 96 well plate assay 50 μL ofpurified human SSAO/VAP-1 (0.25 μg/mL) in 0.1 M NaPO4 buffer (pH 7.4)was added into each well. Test compounds were dissolved in DMSO andtested in a Concentration Response Curve (CRC) with 4-9 data points,typically in the micromolar or nanomolar range after incubation withhuman SSAO/VAP-1 for 30 min at 37° C. After 30 min incubation, 50 μL ofthe reaction mixture containing 600 μM benzylamine (Sigma Aldrich), 120μM Amplex Red (Sigma Aldrich) and 1.5 U/mL horseradish peroxidase (SigmaAldrich) prepared in 0.1 M NaPO4 buffer (pH 7.4) were added to thecorresponding well. The fluorescence unit (RFU) was read every 2.5 minfor 30 min at 37° C. excitation 565 nm and emission 590 (Optima; BMGlabtech). The slope of the kinetics for each well was calculated usingMARS data analysis software (BMG labtech) and this value was used todeduce the IC₅₀ value (Dotmatics). The results are shown in Table 2.

TABLE 2 SSAO/VAP-1, MAO-B and DAO inhibitory activities of examples ofcompounds of the invention and comparative compounds Human SSAO/VAP-1Endogenous Human Human expressed in SSAO/VAP-1 Diamine MAO-B HMEC cellsin rat fat Oxidase Activity IC₅₀ Activity IC₅₀ Activity IC₅₀ ActivityIC₅₀ Compound (micromolar) (nanomolar) (nanomolar) (micromolar) 1 <1<100 <100 <1 2 >1 <100 <100 <0.1 3 >10 <100 <100 >1 4 >0.1 <100 <100 <16 >1 <100 NT <1 7 >10 <100 NT <1 8 >1 <100 NT NT 9 >10 <100 <100 <110 >10 <100 <100 >1 11 >10 <100 <100 >10 13 >0.1 <100 <100 >1 14 >10<100 <100 >10 15 >100 <100 <100 NT 18 >0.1 <100 <100 <0.1 20 >1 <100 NT<1 23 >1 <100 <100 >10 24 >1 <100 <100 >10 25 >1 <100 <100 <1 26 >1 <100NT <1 28 >10 <100 <100 >10 29 >10 <100 <100 >1 30 >10 <100 <100 >1031 >1 <100 <100 <1 32 >10 <100 <100 >10 33 >10 <100 <100 <1 Mofegiline 5nM 19 6 >10

Example 6 Method to Determine the Ability of Compounds of Formula I toInhibit Human Recombinant SSAO/VAP-1 Expressed in HMEC Cells

SSAO/VAP-1 activity was determined using a similar method as describedin Example 5 except for the source of human SSAO/VAP-1.pcDNA-DEST40-hSSAO/VAP-1 was transfected into HMEC cells usinglipofectamine (Invitrogen Ltd). A clone stably expressing humanSSAO/VAP-1 was selected and was stored in liquid nitrogen until celllysate was required for colorimetric assay. Briefly, HMEC cellexpressing human SSAO/VAP-1 were grown in several 10 cm petri dishes,once the cells reached 100% confluency, cells were harvested andhomogenates were prepared. Cells were washed twice with 5 mL of chilledHES buffer (20 mM HEPES, 1 mM EDTA, 250 mM sucrose, pH 7.4). HES buffercontaining 1× protease inhibitor (Sigma Aldrich) and added and cellswere incubated on ice for 3 min. Buffer was removed and cells werescraped and transferred to a centrifuge tube. Cell lysates were preparedby passing through 23 G needle for 10 times and followed by 27 G needlefor 10 times. Alternatively the cell lysates were prepared by using IKAUltra-Turrax T 10 homogenizer for 3 min for every 10 mL of cellsuspensions. Cells were then spun for 5 min at 300×g. The clearsupernatant was transferred to new centrifuge tube and stored at −80° C.until colorimetric assay was performed. Prior to the assay, 0.5 mMpargyline was added in order to inhibit any residue MAO activities. Theassay was performed as described in Example 5. Briefly, 50 μL of celllysate was incubated with test compounds for 30 min at 37° C. Reactionmixtures were added and kinetic was read as described in detail inExample 5. Table 2 shows the data of several compounds of Formula I.

Example 7 Method to Determine the Ability of Compounds of Formula I toInhibit SSAO/VAP-1 in Mouse and Rat Fat Homogenate

Abdominal fat from BALB/c mice, Wistar or Sprague Dawley rats, which aretissues enriched with SSAO/VAP-1—were surgically removed. For every gramof animal abdominal fat tissue, 1 mL of 0.1 M NaPO4 buffer (pH 7.4) wasadded. Tissues were homogenized using IKA Ultra-Turrax T 10 homogenizerfor 3 min, homogenate was centrifuged for 15 min at 3000×g. The middlelayer (clear supernatant) was removed without disturbing the top layer(high fat content) or the debris on the bottom of the tube. SSAO/VAP-1activity was determined by checking the fluorescent signal.K_(m)/V_(max) values were determined and the fat homogenate wasaliquoted and stored at −80 C until assays were performed. Assay wasperformed in a similar fashion as for human SSAO/VAP-1 (Example 5)except, the substrate (benzylamine) concentrations used for mouse fathomogenate and rat fat homogenate were 80 μM and 30 μM respectively. Theresults are shown in Table 2.

Example 8 Method to Determine the Ability of Compounds of Formula I toInhibit Human Recombinant MAO-B

The specificity of invention compounds was tested by determining theirability to inhibit MAO-B activities in vitro. Recombinant human MAO-B(0.06 mg/mL; Sigma Aldrich) was used as source of MAO-B enzymeactivities. The assay was performed in a similar way as for humanSSAO/VAP-1 (Example 5) except, the substrate benzylamine was used at 100μM. Table 2 shows data for several compounds of Formula I.

Example 9 Method to Determine the Ability of Compounds of Formula I toInhibit Human Recombinant Diamine Oxidase

Three human genes are found to encode for copper-containing amineoxidases. Diamine oxidase (DAO) is one of the enzymes produced by theAOC1 gene, named for its substrate preference for diamines. Thespecificity of the compounds of Formula I was tested by determiningtheir ability to inhibit DAO activities in vitro. Recombinant human DAO(2.4 μg/mL) was used as source of DAO enzyme activities. The assay wasperformed as described in the method for human SSAO/VAP-1 (Example 5)except the substrate used was 200 μM putrescine, and the control wellscontained 10 μM aminoguanidine instead of Mofegiline. Table 2 shows datafor several compounds of Formula I.

Example 10 Method to Determine the Ability of Compounds of Formula I toInhibit Lysyl Oxidase

Lysyl oxidase (LOX) is an extracellular copper dependent enzyme whichoxidizes peptidyl lysine and hydroxylysine residues in collagen andlysine residues in elastin to produce peptidylalpha-aminoadipic-delta-semialdehydes. This catalytic reaction can beirreversibly inhibited by β-aminopropionitrile (βAPN) that binds to theactive site of LOX (Tang S. S., Trackman P. C. and Kagan H. M., Reactionof aortic lysyl oxidase with beta-aminoproprionitrile. J. Biol. Chem.1983, 258, 4331-4338). There are five LOX family members; these are LOX,LOXL1, LOXL2, LOXL3 and LOXL4. The specificity of compounds of Formula Iwas tested by determining their ability to inhibit different sources ofLOX family in vitro.

Two sources of enriched LOX were prepared using (1) supernatant fromnormal human lung fibroblast (NHLF) and (2) homogenate from rat skin.Briefly, NHLF was cultured in complete medium containing SingleQuotsupplements with 5% FBS (Lonza Australia Pty Ltd) and FGM-2 medium(Lonza Australia Pty Ltd) in T175 flask until 60% to 80% confluency.Once the optimal confluency was reached, cells were washed twice usingphosphate saline buffer and replaced with medium containing 0.1% FBS andFGM-2 medium. Two to four days later, supernatant was collected andcentrifuged for 5 min at 300×g. Cell debris was removed and LOX proteinswere further enriched using Amicon® Ultra-4 Centrifugal Filter Units,with a 10 kDa cut-off (Millipore Ltd). Briefly, samples were added tothe columns and centrifuged at 4000×g, 4° C. until a final volume of 1mL was obtained. During the centrifugation process, buffer was exchangedusing sodium borate buffer (1.2 M Urea; 0.05 M sodium borate; pH 8.2).Different substrates were tested on the enriched LOX supernatant and thefluorescent signals were measured using colorimetric assay. Thesubstrate specificity and pharmacology properties of the enrichedsupernatant were corroborated with published literatures. The enrichedsupernatant was aliquoted and stored at −80° C.

LOX proteins are found highly expressed on skin (Rucker et al 1995),thus rat skin homogenate were used as a second source for determiningLOX enzyme activities. Briefly, to every gram of finely chopped rat skintissue, 3 mL of phosphate buffered saline was added. Tissues were thenhomogenized using IKA Ultra-Turrax T 10 homogenizer for 3 min. This andall the following homogenizations were performed on ice. The homogenatewas centrifuged (20817×g, 30 min) at 4° C. and the supernatant wasdiscarded. Tissues were resuspended using 4.2M urea-sodium borate bufferand homogenized for approximately 3 min (2.5 mL buffer/g). Homogenatewas incubated overnight at 4° C. Sample was spun (20817×g, 30 min) andsupernatants were collected. Cell pellet underwent two cycles ofhomogenization and the supernatant from each process was collected. Allthe supernatants were pooled and LOX proteins in rat skin homogenatewere enriched using Amicon® Ultra-4 Centrifugal Filter Units, with a 10kDa cut-off. Sample underwent buffer exchange until a concentration of1.2 M urea was reached. Different substrates were tested on the enrichedLOX skin homogenate and the fluorescent signals were measured usingcolorimetric assay. The substrate specificity and pharmacologyproperties were determined. The enriched skin homogenate was aliquotedand stored at −80° C.

The specificity of compounds of Formula I was tested using the twodifferent sources of LOX supernatant from normal human lung fibroblast(NHLF) and homogenate from rat skin. Assays were performed as describedin the method for human SSAO/VAP-1 (Example 5 except these two sourceswere treated with pargyline (0.5 mM), the substrate used was 10 mMputrescine, the control wells contained 10 μM βAPN instead ofMofegiline, and was read at 45° C. Table 2 shows data for severalcompounds of Formula I.

Example 11 Method to Determine the Ability of Compounds of Formula I toInhibit SSAO/VAP-1 when Administered to Mice and Rats

Mice and rats were administered either orally (p.o.) or intravenously(i.v.) with invention compounds at various concentrations ranging from0.1 mg/Kg to 100 mg/Kg. Control group were administered the same volumeof vehicle p.o. or i.v. Abdominal fats, plasma and lung, liver and aortatissue were collected at various time points ranging from 0 to 96 hours.

Each tissue was homogenized in HES buffer with 1× phosphatase inhibitor(Sigma Aldrich) and 1× protease inhibitor (5 mL/g for rats and 20 mL/gfor mice). The homogenate was used to measure SSAO activity as describedin human SSAO/VAP-1 (Example 5), except the mice and rat homogenate wasfurther diluted using 0.1 M NaPO4 buffer (pH 7.4) at 1:5 and 1:20 ratio,respectively. The substrate (benzylamine) concentrations used for mousefat homogenate and rat fat homogenate were 80 μM and 30 μM respectively.The slope of the kinetics for each well was calculated using MARS dataanalysis software. The percentage response was calculated using the SSAOactivity from treated animal tissue normalized to control animals.Graphs were plotted using GraphPad Prism Software. The method describedby Yu, P. H. et al., Involvement of SSAO-mediated deamination in adiposeglucose transport and weight gain in obese diabetic KKay mice, Am JPhysiol Endocrinol Metab 2004, 286: E634-E64 was used to determine thedegree of SSAO/VAP-1 inhibition in plasma. FIGS. 1A-1E, 2A-2E and 3A-3Eshow the dose response profile for Compound 23 in all tissues employingvarious administration protocols.

Example 12 Inhibition of Carrageenan-Induced Rat Paw Edema

Carrageenan-induced paw edema is a widely used test to determine theanti-inflammatory activity of various therapeutic agents and is a usefulexperimental system for assessing the efficacy of compounds to alleviateacute inflammation. Inflammation is induced by intraplantar injection of20 μL of carrageenan suspension (1% in saline) as described (seeRoussin, A. et al., Neutrophil-associated inflammatory responses in ratsare inhibited by phenylarsine oxide. Eur. J. Pharmacol, 1997, 322, 91-96and Wise, L. E. et al., Evaluation of fatty acid amides in thecarrageenan-induced paw edema model. Neuropharmacology, 2008. 54,181-188). Test compound (0.1-100 mg/kg) is given 1 hour prior to theadministration of carrageenan. Paw thickness is measured with electronicdigital calipers prior to and 1, 3, 5, 6 and 24 hours after thecarrageenan injection, to demonstrate greater than 50% inhibition ofedema as compared to control animals.

Example 13 Efficacy in Model of Systemic Inflammation

Evaluation of the efficacy of compounds of the invention is carried outin a model of endotoxemia that consists of intraperitoneal injection ofa high dose of lipopolysaccharisde (LPS) (5 mg/kg) (see Schabbauer, G.et al., PI3K-Akt pathway suppresses coagulation and inflammation inendotoxemic mice. Arterioscler. Thromb. Vasc. Biol., 2004, 24, 1963-1969and Lentsch, A. B. et al., STAT4 and STAT6 regulate systemicinflammation and protect against lethal endotoxemia. J. Clin. Invest.,2001, 108, 1475-1482). Blood samples (50 mL) are collected at 0, 1, 2,4, and 8 hrs after LPS injection and used for blood smears and cytokineevaluation. Plasma concentrations of TNF-α, IL-6, MCP-1 and KC in micetreated with compound (0.1-100 mg/kg) are reduced between 20-80% asmeasured by ELISA. Animal survival rates are recorded for the next 3days and compound treated mice show a 20% greater survival rate.

Example 14 Inhibition of Air Pouch Inflammation in the Mouse

Injection of carrageenan induces inflammation and the pouch serves as areservoir of cells and mediators that can be easily measured in thefluid that accumulates locally.

Animals were anaesthetized and 6 ml of sterile air was injectedsubcutaneously as described (see Romano, M. et al., Carrageenan-inducedacute inflammation in the mouse air pouch synovial model. Role of tumournecrosis factor. Mediators Inflamm, 1997. 6, 32-38). After 3 days thepouches were re-injected with 3 ml of sterile air. On day 6, thecontrols received 1 ml of vehicle; treated controls received 10 mg/kgdexamethasone, and Compound 23 group received 2 mg/kg. 1 hour aftertreatment the mice were injected with 1 ml carrageenan solution into theair pouch. At 4 hours after carrageenan injection, the animals wereeuthanized and the pouches were washed with saline. The exudates wereused for cell count as well as cytokine measurement. Compound 23 treatedmice showed reduced inflammation, with a significant reduction inexudate volume and neutrophil infiltration as well as significantlydiminished TNF-α and IL-6 production (FIG. 4).

Example 15 Inhibition of Leukocyte Migration in CremasterMicrocirculation

The mouse cremaster preparation was used to study the inhibition ofleukocyte migration to the microcirculation and adjacent connectivetissue as described (see Pinho, V. et al., Tissue- andStimulus-Dependent Role of Phosphatidylinositol 3-Kinase Isoforms forNeutrophil Recruitment Induced by Chemoattractants In Vivo. J Immunol2007; 179:7891-7898 and Nanhekhan, L. V., Microcirculatory hemodynamicsof the rat cremaster muscle flap in reduced blood flow states. Ann PlastSurg. 2003 August; 51(2):182-8).

Briefly, an incision was made in the scrotal skin to expose the leftcremaster muscle, which was then carefully removed from the associatedfascia. A lengthwise incision was made on the ventral surface of thecremaster muscle using a cautery. The testicle and the epididymis wereseparated from the underlying muscle and were moved into the abdominalcavity. The muscle was then spread out over an optically clear viewingpedestal and was secured along the edges with a suture. The exposedtissue was superfused with warm bicarbonate-buffered saline. Single,unbranched cremasteric venules (25-40 um in diameter) were selected and,to minimize variability, the same section of cremasteric venule wasobserved throughout the experiment. The number of rolling, adherent, andemigrated leukocytes upon KC or LPS stimulation was determined offlineduring video playback analysis. Rolling leukocytes were defined as thosecells moving at a velocity less than that of erythrocytes within a givenvessel. The flux of rolling cells was measured as the number of rollingcells passing by a given point in the venule per minute. A leukocyte wasconsidered to be adherent if it remained stationary for at least 30 s,and total leukocyte adhesion was quantified as the number of adherentcells within a 100 μm length of venule. Compound 23 (6 mg/kg) was given1 hour prior to the administration of stimulus. Compound 23demonstrated >50% inhibition of rolling and adhesion when compared tothe control group (FIG. 5).

Example 16 Inhibition of Inflammation Upon Induction of the CecalLigation and Perforation (CLP) Insult

The CLP procedure involved a laparotomy and ligation of the cecum,distal to the ileocecal valve as described (see Martin, E. et alPhosphoinositide-3 Kinase γ Activity Contributes to Sepsis and OrganDamage by Altering Neutrophil Recruitment Am. J. Respir. Crit. Care Med.September, 2010 182 (6) 762-773 and Lutterloh, E. C., Inhibition of theRAGE products increases survival in experimental models of severe sepsisand systemic infection. Crit Care. 2007; 11(6):R122).

The cecum was punctured with a needle to induce moderate sepsis;following the puncture a small amount of fecal matter was extruded fromeach puncture. Sham animals received a laparotomy with no manipulationof the cecum. Compound 23 was dosed 6 hours prior to puncture. Followingligation and puncture, the cecum was returned to the abdomen, theperitoneal wall and skin incisions were closed, and the animals wereallowed to recover. Eighteen hours following CLP/sham surgery, aproportion of the animals from each group were sacrificed and the lungswere lavaged. The lavage was centrifuged to isolate inflammatory cellsfor differential cell analysis, while a separate aliquot was used tocount total live cell number using a haemocytometer and lightmicroscopy. Survival was monitored over 7 days. Compared with thevehicle-treated group that showed 50% lethality incidence,compound-treated mice resulted in a statistically significant reductionin lethality with 90% of mice surviving at day 7 (FIG. 6B). In addition,the inhibitory effect of the compound on the inflammatory component ofdisease was seen by reduced total leukocyte in the BALF (FIG. 6A).

Example 17 Inhibition of Chemically Induced Colitis

This procedure is used to screen for compounds which inhibit thedevelopment of colitis as compared to control using the TNBS-inducedcolitis model (see Maslowski, K. M. et al., Regulation of inflammatoryresponses by gut microbiota and chemoattractant receptor GPR43. Nature,2009. 461, 1282-1286). Briefly, mice are sensitised by applying amixture of acetone/olive oil (50:50) with TNBS (50:50, total) on shavedskin between shoulder blades. Seven days later, mice are challengedintra-rectally with 2.5 mg TNBS with 50% ethanol, 3.5 cm from the analverge. Mice are fasted overnight before the intrarectal challenge, andgiven 5% dextrose in the drinking water. Mice are analysed 3 days afterTNBS challenge.

Colitis is also induced by dextran sulphate sodium salt (DSS), asdescribed (see Vieira, A. T. et al., Treatment with a novelchemokine-binding protein or eosinophil lineage-ablation protects micefrom experimental colitis. Am. J. Pathol, 2009. 175. 2382-2891). Micereceive 4% (w/v) DSS in their drinking water ad libitum for 7 days, thenswitch to autoclaved drinking water. Compounds are given throughout theexperimental period at 0.1-100 mg/kg. Mice are sacrificed on the seventhday, and the colon is analysed. For survival studies, mice are followedfor 25 days after start of DSS treatment. Compounds inhibit diseaseprogression as evaluated by less weight loss (20%) and decreasesclinical symptoms. They also delay presence of blood in stools and lossof firmness. Histological analysis of colon sections demonstrate >30%less inflammation. Cytokine measurement shows up to 70% inhibition ofIL5, IL6 and TNFα production.

Example 18 Inhibition of ConA Liver Induced Injury in Mice

Autoimmune liver disease includes autoimmune hepatitis (AIH), a distinctform of acute and chronic inflammatory liver disease in which immunereactions against host antigens have been found to be the majorpathological mechanism. AIH may lead to severe liver disease such asliver cirrhosis. ConA-induced specific liver injury in mice is anexperimental animal model, which has been closely studied in thepathogenesis of the liver injury. T cell mediated immunity and thesubsequent release of TNF-α are considered to play an important role inthis disease.

Concanavalin A (ConA) 10 mg/kg is administered intravenously in saline.Control mice are injected with saline. Transaminase and alkalinephosphatase in blood and liver are >40% reduced by compound at 0.1-100mg/kg. Cytokines, such as IL-6, TNF-α and IL-5, are significantlyreduced, showing up to 75% reduction when compared to control. Hepatichistopathology demonstrates decreased inflammation and tissue damage inthe compound treated group (see Hu, X. D. et al., Preventive effects of1,25-(OH)2VD3 against ConA-induced mouse hepatitis through promotingvitamin D receptor gene expression. Acta Pharmacol. Sin, 2010, 31,703-708; Zhang, X. L. et al., Protective effects of cyclosporine A onT-cell dependent ConA-induced liver injury in Kunming mice. World J.Gastroenterol., 2001, 7, 569-571; Erhardt, A. et al., IL-10, regulatoryT cells, and Kupffer cells mediate tolerance in concanavalin A-inducedliver injury in mice. Hepatology, 2007, 475-485).

Example 19 Inhibition of Parkinson's Disease Pathology in Rats

Model A: Systemic Exposure to LPS to Promote Neurodegeneration

Parkinson's disease is a pathological, age-related neurodegenerativedisorder, characterized by a specific and progressive degeneration ofdopaminergic neurons. Peripheral exposure to LPS, a potent inducer ofinflammation in rodents, has been shown to result in neuroinflammation,persistent microglial activation, delayed and progressive dopamineneurons loss in the substantia nigra, similar to that observed inParkinson's Disease. Recent evidence has implicated inflammation in theneurodegeneration of nigrostriatal dopaminergic neurons, and LPS wasshown to promote it (see Qin, L. et al. Systemic LPS causes chronicneuroinflammation and progressive neurodegeneration, 2007 Glia,453-462).

Long Evans rats were dosed intraperitoneal (ip) with 2 mg/kg of Compound9 or vehicle 1 h before the first (time 0 h) and the third (time 24 h)injections of LPS. At time 0 the animals received a dose of 10 mg/kg ofLPS. At time 6 and 24 h the animals were dosed with 3 mg/kg of LPSsolution, ip. 30 h after the first LPS injection, the animals receivedip injections of lethabarb and were transcardially perfused with 400 mlPBS at 4° C. followed by 400 ml of 4% paraformaldehyde (PFA). The brainswere post fixed overnight in 4% PFA at 4° C. followed by 20% sucrosesolution for 24 h. 30 μm sections were collected and stained forimmunofluorescence, immunohistochemistry and western blot analysis. Thegroup treated with Compound 9 showed reduced neutrophil infiltration inthe dorso-lateral striatum and hippocampus, and a reduction ofmicroglial cell recruitment and activation (dendrites length, surfaceand volume) in the substantia nigra and dorso-lateral striatum (FIG. 7).

Model B: Localized Exposure to LPS to Promote Neurodegeneration

Direct injection of LPS in selected areas of the brain can be performedin order to induce a localized inflammatory response in the brain. Thedopaminergic neurons are more vulnerable to inflammation basedneurotoxicity, and the local LPS injections in relevant areas such assubstantia nigra and striatum have been used as a model for Parkinson'sDisease (see Liu, M., & Bing, G. Lipopolysaccharide animal models forParkinson's disease. Parkinson's disease, 2011, 327089; Choi, D.-Y. etal. Striatal neuroinflammation promotes Parkinsonism in rats. PloS one,2009, 4(5), e5482). LPS has also been shown to promote nigraldopaminergic neuron degeneration (see Machado, A. et al., Inflammatoryanimal model for Parkinson's Disease: The intranigral injection of LPSinduced the inflammatory process along with the selective degenerationof nigrostriatal dopaminergic neurons. ISRN Neurology, 2011, 1-16).

A solution containing 2 μL of 1 mg/mL of LPS is injected in the leftsubstantia nigra of female rats previously anesthetized. Animals aretreated with 0.1-100 mg/Kg of compound and the results show up to 80%decreases in inflammation with less activation of microglia as comparedto control animals. Vehicle treated animals are accompanied by loss ofdopaminergic neurons and decreases of the intracellular content ofdopamine (DA), effects which are significantly inhibited by thecompound. The average loss of the dopaminergic system in the vehicletreated groups is around 35%, whilst in the compound treated group it is<20%.

Example 20 Inhibition of Inflammation Associated with Stroke in Mice

The development of the brain tissue damage in stroke is composed of animmediate component followed by an inflammatory response with secondarytissue damage after reperfusion. The ischemia/reperfusion model mimicsthe tissue damage as well as inflammatory component (see Hase, Y. etal., Cilostazol, a phosphodiesterase inhibitor, prevents no-reflow andhaemorrhage in mice with focal cerebral ischemia. Exp. Neurol., 2012,233(1), 523). Mice are subjected to middle cerebral arteryocclusion/reperfusion surgery by introducing a nylon monofilament intothe right common carotid artery (CCA). It is carefully advanced to 11 mmfrom the carotid artery bifurcation and a proximal occlusion of theright middle cerebral artery is established. After 90 min occlusion, thefilament is withdrawn to allow reperfusion for another 22.5 hr. Animalsare treated with compound 0.1-100 mg/Kg and show up to 50% reduction inplatelet aggregation and leukocyte plugging in the micro vessels.Treatment significantly reduces mortality rate with >80% of animalsurvival.

Example 21 Inhibition of Acute Lung Inflammation in the LPS Driven Model

Inflammation was induced by instillation of LPS into the lungs of miceusing an tracheal surgery challenge method (see Innate immune responsesto LPS in mouse lung are suppressed and reversed by neutralization ofGM-CSF via repression of TLR-4. Am. J. Physiol. Lung Cell. Mol.Physiol., 2004, L877-85; and Harrod, K. S., A. D. Mounday, and J. A.Whitsett, Adenoviral E3-14.7K protein in LPS-induced lung inflammation.Am. J. Physiol. Lung Cell. Mol. Physiol., 2000, 278, L631-9). Briefly, 1hour after treatment with 10 mg/kg of Dexamethasone or 2 mg/kg ofCompound 9, mice were anesthetized, a midline incision was made in theneck, the muscle layers separated by blunt dissection, and 1 ml/kg LPS(20 mg/kg) or vehicle injected into the trachea. The incision was closedwith wound clips and the mice returned to cages.

Six hours after LPS/saline injection, the mice were anesthetized, thewound clips removed, the trachea was cannulated with a 23 G bluntneedle, and the lungs lavaged eight times with 0.5 ml heparinizedsaline. The lavage was pooled, gently inverted, and a sample retainedfor white blood cell (WBC) differential analysis. The remainder of thelavage was centrifuged, the supernatants used for cytokine analysis.Compound 9 showed a significant reduction in neutrophil infiltration anda diminution of IL-6 and TNF-α levels compared to controls (FIG. 8).

Example 22 Inhibition of Lung Allergic Inflammation of Viral InfectedMice

Early-life respiratory viral infections, notably with respiratorysyncytial virus (RSV), increase the risk of subsequent development ofchildhood asthma. Infection with pneumonia virus of mice (PVM), whichbelongs to the same family (Paramyxoviridae) and genus (Pneumovirus) asRSV, provides a model of RSV disease (see Rosenberg, H. F. et al., Thepneumonia virus of mice infection model for severe respiratory syncytialvirus infection: identifying novel targets for therapeutic intervention.Pharmacol. Ther., 2005, 105, 1-6). Allergic airway inflammation,including recruitment of eosinophils, is prominent in animals that areneonatally infected with PVM and then challenged with OVA antigen (seeSiegle, J. S. et al., Early-life viral infection and allergen exposureinteract to induce an asthmatic phenotype in mice. Respir. Res., 2010,11, 14).

On both days 1 and 2 of life, mice are intranasally inoculated with 2pfu (PVM J 3666 strain ˜1×10⁵ pfu/mL) in 5 μL phosphate buffered saline(PBS) on the external nares. Control animals are sham-infected with PBSalone. Intranasal sensitisation to OVA is performed either at days 1 and2 of life or at days 28 and 29, with 5 μg OVA/5 μL PBS or 100 μg/40 μLrespectively. Mice receive low-level aerosol challenge with ovalbumin(mass concentration of ≈3 mg/m3 of ovalbumin for 30 min/day, 3 days/weekfor 4 weeks). This is followed by a single moderate-level challenge (≈30mg/m³ for 30 minutes) to induce the changes of an acute exacerbation.The purpose of this study is to assess anti-inflammatory effect of thecompound (0.1-100 mg/kg) in mice that are predisposed to the developmentof features of asthma due to early-life infection.

Bronchoalveolar lavage (BAL) is performed for recovery of airway luminalcells. This procedure is achieved by intratracheal instillation of 800μL of PBS/mouse. The total number of leukocytes is counted using ahaemocytometer. Cytospin slides are prepared from BAL fluid and thenstained with Wright-Giemsa stain for differential cell count. Cells areclassified into mononuclear cells, eosinophils, neutrophils andlymphocytes according to standard morphologic criteria and at least 200cells were counted per slide under light microscopy. For lung histology,lungs are perfused, inflated and fixed in 10% buffered formalin beforeimmunohistochemichal analysis. The extent of the leukocyte infiltrate isscored as 0, minimal or no inflammation; 1, mild inflammation, onlyperivascular or peribronchiolar; 2, moderate inflammation, someparenchymal involvement; 3, marked inflammation, widespread parenchymalinvolvement; 4, severe inflammation as previously described. Compoundsare administered at 0.1 mg/kg-100 mg/kg and animals show a reduction of40-80% in neutrophil infiltration, diminution of IL-6 and TNFα of up to30% compared to controls.

Example 23 Inhibition of Exacerbation in an HDM-Induced Asthma Model

Respiratory infections, which are predominantly caused by rhinovirus inpeople with asthma, exacerbate airway inflammation and furthercontribute to disease burden and healthcare cost. The rhinovirusexacerbated house dust mite (HDM) model was used to study the effect ofCompound 23 in a model of allergic asthma (Collison, A. et al. The E3ubiquitin ligase midline 1 promotes allergen and rhinovirus-inducedasthma by inhibiting protein phosphatase 2A activity. Nat. Med. 2013,19(2): 232-7).

Mice were sensitized and challenged by exposing them intranasally tocrude HDM extract (50 μg daily at days 0, 1 and 2 followed by fourexposures of 5 μg HDM daily from day 14 to day 17 delivered in 50 μl ofsterile saline). Animals were infected (day 18, 1 d after last HDMextract challenge) with 50 μl infective or ultraviolet light(UV)-inactivated RV1B41 (2.5×106 median tissue culture infective dose)intranasally. Compounds were dosed at 0.1-100 mg/kg 1 hour prior torhinovirus challenge. Mice were killed 24 h after the last allergen orrhinovirus challenge. Cytospin slides were prepared from Bronchoalveolarlavage fluid and then stained with Wright-Giemsa stain for differentialcell count. Cells are classified into mononuclear cells, eosinophils,neutrophils and lymphocytes according to standard morphologic criteriaand at least 200 cells were counted per slide under light microscopy.Animals treated with Compound 23 at 6 mg/kg showed a significantreduction in neutrophil infiltrate in the BALF (FIG. 9A) and reducedairway hyper reactivity in response to metacholine challenge back tothat of the control group (FIG. 9B).

Example 24 Inhibition of Cutaneous Inflammation in the SCID Mouse Modelof Psoriasis

Psoriasis is a common inflammatory skin disease characterized byabnormal epithelial differentiation, extensive capillary formation inthe papillary dermis, and accumulation of inflammatory leukocytesincluding T lymphocytes, NK lymphocytes, and granulocytes.Transplantation of human skin onto immunocompromised mice (severecombined immunodeficiency [SCID] mice) provides a model to studypsoriasis. Using this approach, epidermal thickening, extensive rete pegformation, and presence of inflammatory cells are maintained for anextended period in the transplanted skin (see Zeigler, M. et al.,Anti-CD11a ameliorates disease in the human psoriatic skin-SCID mousetransplant model: comparison of antibody to CD11a with Cyclosporin A andclobetasol propionate. Lab. Invest, 2001, 81, 1253-1261 and Nickoloff,B. J. et al., Severe combined immunodeficiency mouse and human psoriaticskin chimeras. Validation of a new animal model. Am. J. Pathol., 1995,146, 580-588).

SCID mice (6-8 weeks old) are prepared for orthotopical skin xenografts.Human skin xenografts (measuring 1.5×1.5×0.05 cm) are sutured to theflank area of each SCID mouse with absorbable Dexon suture. Dressingsare changed every 2 days, and animals are maintained pathogen-freethroughout the study. Human skin/SCID mice chimeras are sacrificed at 4or 6 weeks after xenograft transplantation (as this period of timeassured adequate acceptance and healing). Xenograft biopsies areprocessed for cytokine ELISA as well as histopathology analysis. Aftertransplantation, compound treated group (0.1-100 mg/kg) show a 20-50%reduction in inflammation in the dermis and epidermis, compared with thevehicle-treated group. In addition, cytokines such as IL-6 and TNFα areinhibited by up to 80% by compound treatment.

Example 25 Antimicrobial Activity—Klebsiella pneumoniae Infection

The efficacy of the compound was investigated in a model of pulmonaryinfection caused by the Gram-negative bacterium Klebsiella pneumoniae.The outcomes were the differences between compound and control inlethality rates, bacterial counts and inflammatory indices followingpulmonary infection of mice (see Soares, A. C. et al., Dual function ofthe long pentraxin PTX3 in resistance against pulmonary infection withKlebsiella pneumoniae in transgenic mice. Microbes Infect., 2006, 8,1321-1329.).

BALB/c mice (8 weeks old) were divided in 3 groups; 2 infected and 1uninfected. Infected groups: Group A, animals were administered vehicleorally; Group B, animals were administered 2 mg/kg of compound orally;and Group C animals were uninfected. Broncheoalveolar lavage fluid(BALF) was collected to determine total number of leukocytes. Cytospinslides were prepared from BAL fluid and then stained with Wright-Giemsastain for differential cell count. Cells are classified into mononuclearcells, eosinophils, neutrophils and lymphocytes according to standardmorphologic criteria and at least 200 cells were counted per slide underlight microscopy. For bacterial counts, lung was homogenised, seriallydiluted and plated on MacConkey agar plates. Colony forming units werecounted at the end of 24 hours incubation at 37° C. Animal survivalrates were recorded for the next 10 days.

Compared with the vehicle-treated group that showed a 45% lethalityincidence, Compound 23 treated mice showed a statistically significantreduction in lethality with 100% of mice surviving (p=0.0597) after 8days (FIG. 10A). In addition, the inhibitory effect of Compound 23 onthe inflammatory component of disease was seen in reduced leukocyteinfiltrate to the BALF (FIG. 10B).

Example 26 Inhibition of Chronic Obstructive Pulmonary Disease

Chronic Obstructive Pulmonary Disease (COPD) is a debilitating disorderof the lung. The disease is characterized by chronic airwayinflammation, mucus hypersecretion, airway remodeling, and emphysema,which lead to reduced lung function and breathlessness. Airflowlimitation is usually both progressive and associated with an abnormalinflammatory response of the lungs to noxious gases and particles.Cigarette smoke elicits a repetitive inflammatory insult that isbelieved to, through the actions of mediators such as proteinases, leadto structural and functional changes in the lung. Moreover, patientswith COPD are more susceptible to respiratory tract infections (seeBeckett, E. L., A new short-term mouse model of chronic obstructivepulmonary disease identifies a role for mast cell tryptase inpathogenesis. J Allergy Clin Immunol. 2013 March; 131(3):752-762.e7;Guerassimov, A., The Development of Emphysema in Cigarette Smoke-exposedMice Is Strain Dependent. Am. J. Respir. Crit. Care Med. November, 2004(170) 974-980 and Morris, A., Comparison of Cigarette Smoke-InducedAcute Inflammation in Multiple Strains of Mice and the Effect of aMatrix Metalloproteinase Inhibitor on These Responses. JPET December2008 (327) 851-862).

BALB/c mice were simultaneously exposed to cigarette smoke (twelve 3R4Freference cigarettes [University of Kentucky, Lexington, Ky] twice perday and 5 times per week for 1 to 12 weeks) by using a custom-designedand purpose-built nose-only, directed-flow inhalation and smoke-exposuresystem (CH Technologies, Westwood, N.J.) housed in a fume and laminarflow hood. Each exposure lasted 75 minutes. Nose-only exposure wasachieved by using specialized containment tubes that delivered smoke andnormal air directly to the animal's nose. This protocol allowed a moreintensive delivery of smoke than whole-body exposure systems. For thefirst 2 days, mice were exposed to 1 session of smoking with 12 puffsfrom each cigarette to allow acclimatization. Smoke was delivered in2-second puffs, with 30 seconds of normal air between each puff. Afterday 2, the mice were subjected to 2 sessions in which they were exposedto the smoke from 12 cigarettes (morning and afternoon, separated by arecovery period). Compound 23 was given at 2 mg/kg from week 6 onwardsof the experimental procedure and significantly inhibited lung collagencontent (FIG. 11).

Example 27 Inhibition of CCl₄ Induced Liver Fibrosis

An analysis of the use of VAP-1/SSAO inhibitors to treatinflammatory/fibrotic diseases is performed through the use of a CCl₄induced liver fibrosis model. Liver injury is frequently followed bycomplete parenchymal regeneration due to regenerative potency ofhepatocytes. However, the concomitant activation of fat-storing cellsleads to extracellular matrix accumulation accompanied by recurrenthepatocyte necrosis, inflammation, and regenerative processes, andcauses liver fibrosis and consequently liver cirrhosis (see Natsume, M.et al., Attenuated liver fibrosis and depressed serum albumin levels incarbon tetrachloride-treated IL-6-deficient mice. J. Leukoc. Biol.,1999, 66, 601-608.).

Liver fibrosis in the male Sprague Dawley (SD) rats was induced by oralapplication of CCl₄ (2.5 μL/g of CCl₄ olive solution, 3 times a week).Vehicle (PBS), and the positive control imatinib mesylate (2.5 mg/kg)were given to the rats from day 1 to day 28, and Compound 23 (6 mg/kg)was given to the rats from day 14 to day 28. Compound 23 demonstrated aclear trend of decreased levels of fibrotic tissue, as represented by adecrease in Sirius red staining (FIG. 12C). Moreover, Compound 23 showedliver function protective effects and a reduction in inflammation whichwere evidenced by significantly decreased levels of serum ALT and AST(FIGS. 12A & 12B) and a reduction in inflammatory score (12D) whencompared to the CCl₄ only group.

Example 28 Inhibition of Non-Alcoholic Steatohepatitis (NASH) InducedLiver Fibrosis

An analysis of the use of VAP-1/SSAO inhibitors to treatinflammatory/fibrotic diseases is performed through the use of anon-alcoholic steatohepatitis (NASH) induced liver fibrosis model. STAMmodel of NASH was induced in 30 male mice by a single subcutaneousinjection of streptozotocin solution 2 days after birth and feeding withhigh fat diet (HFD, 57 kcal % fat) after 4 weeks of age to 10 weeks ofage. From 7 weeks of age mice were orally administered daily dose ofvehicle (PBS), Compound 23 (6 mg/kg) or the positive control Telmisartan(10 mg/kg) for 3 weeks. Compound 23 reduced both inflammation andnon-alcoholic fatty liver disease (NAFLD) scores upon clinicalexamination (FIGS. 13A & 13B). Fibrosis, as evidenced by a reduction ofSirius red-positive area (FIG. 13C) was also reduced.

Example 29 Inhibition of Uveitis

This procedure is to determine inhibition of uveitis by compound(s)according to the invention. Uveitis is a complex inflammatory eyedisease that can lead to blindness. It can affect any part of the eyeand is characterized by the accumulation of leukocytes in oculartissues. Current therapies for uveitis include corticosteroids andchemotherapeutic agents to reduce inflammation. However, the grave sideeffects of these drugs, such as increased intraocular pressure orcytotoxicity limit their use (see Moorthy, R. S. et al., Glaucomaassociated with uveitis. Surv. Ophthalmol., 1997, 41, 361-394 andLightman, S., New therapeutic options in uveitis. Eye 1997, 11,222-226).

Thirty (30) Lewis albino rats were divided into four (4) groups. Forthree groups out of 4, ocular inflammation was induced by a singlefootpad injection of 1 mg/kg lipopolysaccharide (LPS from SalmonellaTyphimurium). Compound 23 (2 mg/kg) and vehicle were administered byoral gavage (1 ml/kg) 1 hour before induction (Day 0). Reference item(dexamethasone, 2 mg/kg) was administered by intravenous injection (2.5ml/kg) just after induction (Day 0). Ocular inflammation was assessed byclinical examination and quantification of neutrophils, eosinophils andproteins in aqueous humor, 24 h after induction.

Clinical Examination of Inflammation; Animals were examined with aslit-lamp at baseline (Day −1) then 24 h after induction (Day 1). Theinflammation in each animal was graded using a scoring system asdescribed (Devos A. et al., Systemic antitumor necrosis factor antibodytreatment exacerbates Endotoxin Induced Uveitis in the rat. Exp. Eye.Res. 1995; 61: 667-675.). Flare, miosis and hypopion were scored forabsence (0), or presence (1), iris hyperemia and cells in the anteriorchamber were scored for absence (0), or mild (1) or severe presence (2).The maximum score (sum of the five parameter scores) is 7. In the grouptreated with Compound 23, a 33% reduction in the severity of the ocularinflammation, compared with the score observed for the vehicle group,was detected 24 hours after induction and 25 hours after oraladministration (FIG. 14A).

At the end of the clinical evaluation (24 h after induction), animalswere anesthetized by an intramuscular injection of a mixed solution ofRompun® (xylazine) and Imalgene® 1000 (ketamine) and euthanized bycardiac injection of overdosed pentobarbital. The aqueous humor wascollected immediately for each eye.

Quantification of Cellular Infiltration in Aqueous Humor; Infiltratedneutrophils and eosinophils were manually counted in cytologicalpreparation of aqueous humor samples diluted 10-fold with PBS beforeGiemsa staining. A significant diminution in eosinophils (mean±SEM:8.9±1.7 cells/μL, n=20) was observed for the group treated with Compound23 versus the group treated with the vehicle (p=0.033) (FIG. 14B).

Example 30 Inhibition of Macular Degeneration

Age-related macular degeneration (AMD) is the leading cause of blindnessand occurs in two major forms. The first is a geographic atrophy (‘dry’)form that is defined by degeneration of photoreceptors and the retinalpigmented epithelium (RPE) near the macula, the accumulation oflipofuscin (A2E), and the formation of drusen. The second is a ‘wet’form that is associated with choroidal neovascularization (see Randazzo,J. et al., Orally active multi-functional antioxidants areneuroprotective in a rat model of light-induced retinal damage. PLoSOne, 2011, 6 e21926 and Davis, S. J. et al., The Effect of Nicotine onAnti-Vascular Endothelial Growth Factor Therapy in a Mouse Model ofNeovascular Age-Related Macular Degeneration. Retina, 2011).

Model A: Light Model

After two weeks of dark adaptation, rats from each group are exposed todamaging light for three hours to 1000 1× of cool white fluorescentlight (light-damaged rats, LD). The control rats in each group are alsoplaced into the light box apparatus for three hours, but not exposed tolight (non-light-damaged rats, NLD). Oxidative stress markers wereevaluated immediately after light exposure. Compound treated animals0.1-100 mg/kg show >20% reduction in oxidative stress as seen byevaluation of neural retinas, which are dissected—followingeuthanasia—from the enucleated eye. For functional and morphologicalassessment, rats are returned to the dark environment after exposure andretinal function is assessed by ERG, 5 to 7 days later. Following ERGanalysis, the rats are euthanized and the enucleated eyes areimmediately processed for quantitative morphology. Compound treatedgroup demonstrate a reduction in severity of disease as seen bydecreases in morphological changes of the eyes as compares to controlanimals.

Model B: Laser Model

CNV is induced by laser photocoagulation in mice with an argon laser(spot size, 50 mm; duration, 0.05 seconds; power, 260 mW). Three laserspots are placed in each eye close to the optic nerve. Production of avaporization bubble at the time of laser confirms the rupture of BM.Animals from each group are sacrificed on days 1, 3, 5, and 7post-laser. Compared with control, the compound-treated mice (0.1-100mg/kg) show a significant reduction in size (by 20%) and incidence ofCNV (>40%) as determined by microscopy.

Example 31 Inhibition of Cancer Progression

B16F10 melanoma cells (4×10⁵ cells/animal) are injected in the shavedadnominal region of the animal as described in Marttila-Ichihara, F. etal., Small-Molecule Inhibitors of Vascular Adhesion Protein-1 Reduce theAccumulation of Myeloid Cells into Tumors and Attenuate Tumor Growth inMice. The Journal of Immunology, 2010, 184, 3164-3173. The growth of thetumor is followed by measuring the dimensions using electroniccallipers. Tumor progression is diminished in compound treated animals(0-1.-100 mg/kg), with up to 25% less tumor growth when compared tocontrol group. Compound treated groups show attenuated myeloid cellaccumulation in the tumors, showing >40% less cell infiltration; inaddition treated mice demonstrate inhibited neoangiogenesis.

All patents and other references cited in the specification areindicative of the level of skill of those skilled in the art to whichthe invention pertains, and are incorporated by reference in theirentireties, including any tables and figures, to the same extent as ifeach reference had been incorporated by reference in its entiretyindividually.

One skilled in the art would readily appreciate that the presentinvention is well adapted to obtain the ends and advantages mentioned,as well as those inherent therein. The methods, variances, andcompositions described herein as presently representative of preferredembodiments are exemplary and are not intended as limitations on thescope of the invention. Changes therein and other uses will occur tothose skilled in the art, which are encompassed within the spirit of theinvention, are defined by the scope of the claims.

It will be readily apparent to one skilled in the art that varyingsubstitutions and modifications may be made to the invention disclosedherein without departing from the scope and spirit of the invention. Forexample, variations can be made to provide additional compounds ofFormula I and/or various methods of administration can be used. Thus,such additional embodiments are within the scope of the presentinvention and the following claims.

The invention illustratively described herein suitably may be practicedin the absence of any element or elements, limitation or limitationswhich is not specifically disclosed herein. The terms and expressionswhich have been employed are used as terms of description and not oflimitation, and there is no intention that in the use of such terms andexpressions of excluding any equivalents of the features shown anddescribed or portions thereof, but it is recognized that variousmodifications are possible within the scope of the invention claimed.Thus, it should be understood that although the present invention hasbeen specifically disclosed by preferred embodiments and optionalfeatures, modification and variation of the concepts herein disclosedmay be resorted to by those skilled in the art, and that suchmodifications and variations are considered to be within the scope ofthis invention as defined by the appended claims.

In addition, where features or aspects of the invention are described interms of Markush groups or other grouping of alternatives, those skilledin the art will recognize that the invention is also thereby describedin terms of any individual member or subgroup of members of the Markushgroup or other group.

Also, unless indicated to the contrary, where various numerical valuesare provided for embodiments, additional embodiments are described bytaking any 2 different values as the endpoints of a range. Such rangesare also within the scope of the described invention.

Thus, additional embodiments are within the scope of the invention andwithin the following claims.

The invention claimed is:
 1. A compound of Formula II:

or a pharmaceutically acceptable salt; wherein: R⁵ is an optionallysubstituted arylene group; R⁶ is selected from

R⁷ and R⁸ are independently selected from the group consisting ofhydrogen, optionally substituted C₁₋₆alkyl and optionally substitutedC₃₋₇cycloalkyl; and X is CH₂, oxygen, sulfur or SO₂; and wherein theterm “arylene” refers to phenylene or naphthylene; and wherein the term“optionally substituted” means the group to which this term refers maybe unsubstituted, or may be substituted with one or more groupsindependently selected from halogen, C₁-C₆alkyl, C₂-C₆alkenyl,C₁-C₆haloalkyl, C₁-C₆alkoxy, hydroxy(C₁₋₆)alkyl, C₃-C₆cycloalkyl, C(O)H,C(O)OH, NHC(O)H, NHC(O)C₁-C₄alkyl, C(O)C₁-C₄alkyl, NH₂, NHC₁-C₄alkyl,N(C₁-C₄alkyl)₂, NO₂, OH and CN.
 2. The compound of claim 1, wherein R⁵is an unsubstituted phenylene group or a phenylene group substituted byone or more groups independently selected from alkyl, halo, alkoxy andhaloalkyl, preferably independently selected from methyl, fluorine,chlorine, bromine, OCH₃ and CF₃.
 3. The compound of claim 1, wherein Xis oxygen.
 4. The compound of claim 1 wherein R⁷ and R⁸ areindependently selected from the group consisting of hydrogen andC₁₋₆alkyl.
 5. The compound of claim 1 wherein R⁶ is


6. The compound of claim 1 wherein R⁶ is


7. The compound of claim 1 wherein said compound is selected from thegroup consisting of:  3

(E)-4-(2-(Aminomethyl)-3- fluoroallyloxy)benzamide; 11

(E)-4-(2-(Aminomethyl)-3- fluoroallyloxy)benzene- sulfonamide; 13

(E)-4-(2-(Aminomethyl)-3- fluoroallyloxy)-N,N- dimethylbenzamide; 14

(E)-4-(2-(Aminomethyl)-3- fluoroallyloxy)-N,N-dimethylbenzenesulfonamide; 17

(E)-4-(2-(Aminomethyl)-3- fluoroallyloxy)-N-tert-butylbenzenesulfonamide; 19

(E)-4-(2-(Aminomethyl)-3- fluoroallyloxy)-N-tert-butyl-3-fluorobenzamide; 21

(E)-4-(2-(Aminomethyl)-3- fluoroallyloxy)-N-tert-butyl-2-(trifluoromethyl)benzamide; 23

(E)-4-(2-(Aminomethyl)-3- fluoroallyloxy)-N-tert- butylbenzamide; 24

(E)-4-(2-(Aminomethyl)-3- fluoroallyloxy)-N,N- diethylbenzamide; 25

(E)-4-(2-(Aminomethyl)-3- fluoroallyloxy)-N- methylbenzamide; 28

(E)-4-(2-(Aminomethyl)-3- fluoroallyloxy)-N- methylbenzenesulfonamide;30

(E)-4-(2-(Aminomethyl)-3- fluoroallyloxy)-N- ethylbenzenesulfonamide; 32

(E)-4-(2-(Aminomethyl)-3- fluoroallyloxy)-N-isopropylbenzenesulfonamide; 34

(Z)-4-(3-(Aminomethyl)-4- fluorobut-3-enyl)-N-tert- butylbenzamide; 39

(E)-4-(2-(Aminomethyl)-3- fluoroallyloxy)-N- isopropylbenzamide;

or a pharmaceutically acceptable salt thereof.
 8. The compound of claim1 wherein said compound is selected from the group consisting of: 13

(E)-4-(2-(Aminomethyl)-3- fluoroallyloxy)-N,N- dimethylbenzamide; 23

(E)-4-(2-(Aminomethyl)-3- fluoroallyloxy)-N-tert- butylbenzamide; 24

(E)-4-(2-(Aminomethyl)-3- fluoroallyloxy)-N,N- diethylbenzamide; 25

(E)-4-(2-(Aminomethyl)-3- fluoroallyloxy)-N- methylbenzamide; 39

(E)-4-(2-(Aminomethyl)-3- fluoroallyloxy)-N- isopropylbenzamide;

or a pharmaceutically acceptable salt thereof.
 9. The compound of claim1 wherein said compound is(E)-4-(2-(aminomethyl)-3-fluoroallyloxy)-N-tert-butylbenzamide or(E)-4-(2-(aminomethyl)-3-fluoroallyloxy)-N-tert-butylbenzamidehydrochloride.
 10. A pharmaceutically acceptable salt of a compoundaccording to claim
 1. 11. The pharmaceutically acceptable salt accordingto claim 10 characterized in that it is an acid addition salt.
 12. Thepharmaceutically acceptable salt according to claim 11 characterized inthat the acid addition salt is selected from the group consisting ofhydrochlorides, hydrobromides, sulfates, formates, acetates, lactates,malates, tartrates, citrates, ascorbates, succinates, maleates,butyrates, valerates and fumarates.
 13. The pharmaceutically acceptablesalt according to claim 11 characterized in that the acid addition saltis a hydrochloride salt.
 14. A composition comprising a compoundaccording to claim 1, or a pharmaceutically acceptable salt, and atleast one pharmaceutically acceptable excipient, carrier or diluent. 15.A method of inhibiting the amine oxidase activity ofsemicarbazide-sensitive amine oxidase/vascular adhesion protein-1(SSAO/VAP-1) in a subject in need thereof, said method comprisingadministering to said subject an effective amount of a compoundaccording to claim 1, or a pharmaceutically acceptable salt thereof, toeffect a positive therapeutic response.
 16. A method of therapeuticallytreating a disease associated with or modulated bysemicarbazide-sensitive amine oxidase/vascular adhesion protein-1(SSAO/VAP-1), said method comprising administering to a subject in needthereof a therapeutically effective amount of a compound according toclaim 1, or a pharmaceutically acceptable salt thereof.
 17. The methodof claim 16 wherein the disease is inflammation.
 18. The method of claim17 wherein said inflammation is associated with liver disease.
 19. Themethod of claim 17 wherein said inflammation is associated withrespiratory disease.
 20. The method of claim 17 wherein saidinflammation is associated with cystic fibrosis.
 21. The method of claim17 wherein said inflammation is associated with asthma or chronicobstructive pulmonary disease.
 22. The method of claim 17 wherein saidinflammation is associated with ocular disease.
 23. The method of claim16 wherein the disease is a diabetes-induced disease selected from thegroup consisting of diabetic nephropathy, glomerulosclerosis, diabeticretinopathy, non-alcoholic fatty liver disease and choroidalneovascularisation.
 24. The method of claim 16 wherein the disease is aneuroinflammatory disease.
 25. The method of claim 16 wherein thedisease is selected from the group consisting of liver fibrosis, livercirrhosis, kidney fibrosis, idiopathic pulmonary fibrosis andradiation-induced fibrosis.
 26. The method according to claim 16characterized in that the disease is non-alcoholic steatohepatitis(NASH).
 27. The method according to claim 23 characterized in that thedisease is diabetic retinopathy.
 28. The method of claim 16 wherein thedisease is cancer.